PONE-D-24-17432

Ptgds downregulation protect Vestibular hair cells from aminoglycoside-induced vestibulotoxicity

PLOS ONE

Dear Dr.  Nie,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

==============================

ACADEMIC EDITOR:

This manuscript focused the PTGDs in vestibulotoxicity-induced vestibular hair cell. 

We have deeply review your manuscript and decided to major revision.

Thank you.

==============================

Please submit your revised manuscript by Jul 19 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

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We look forward to receiving your revised manuscript.

Kind regards,

Sujeong Jang

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

Reviewer #3: Partly

Reviewer #4: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: No

Reviewer #4: Yes

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors of this article hope to study the mechanism of aminoglycoside ototoxicity, which has certain research value, but there are still the following questions:

1. There are many types of aminoglycosides. Why choose neomycin for research? Should more drugs be added for research?

2. There are many differentially expressed genes screened in the neomycin injury in vitro model. Why choose Ptgds as the research gene?

3. Ptgds is a downregulated gene screened in the neomycin injury in vitro model. Why do we need to knock out and silence it in subsequent experiments?

4. In addition to gene knockout, should we increase the study of overexpression?

Reviewer #2: The manuscript by Chen Chen and colleagues, entitled " Ptgds downregulation protect vestibular hair cells from aminoglycoside-induced vestibulotoxicity", is a study investigating the role of the Ptgds in the organ of vestibular hair cell protection.

This study demonstrated the significant upregulation of Ptgds in utricle explants and HEI-OC1 cells in response to neomycin-induced injury. And the knockdown of Ptgds by using siRNA effectively protected and attenuated neomycin-induced vestibular hair cells loss. In vitro experiment confirmed that Ptgds silencing was sufficient to mitigate neomycin-induced ROS production and apoptotic death in hair cells through the inhibition of inflammatory related genes, such as COX2, IL6 and iNOS. The logic of this article is clear, the evidence chain supporting the conclusion is sufficient, and it is innovative.

There are some issues and small comments as follows:

• In the paragraph 2 of Introduction section: “While prior work conducted by our team has revealed the overexpression of Ptgds in vestibular inner ear hair cells in response to neomycin injury…….”. Relevant references should be added here. Otherwise, this manner of expression would not be very appropriate. The same issue exists in the second paragraph of the discussion section.

• In the section of 2.5: What’s ECL stand for?

• In the section of 2.6: What does 1x103/well mean? And what is the siRNA sequence of sense strand?

• In the statistical analysis section, the description should include the analysis software used and the statistical methods employed, such as t-tests, analysis of variance (ANOVA), etc.

• In the last sentence of the first paragraph of the section 3.1, it is mentioned that the expression of Ptgds is significantly downregulated after neomycin injury. This is inconsistent with the results in other parts of the article. Please verify！

• In the first paragraph of section 3.2: “The viability of cells in the control, neomycin-treated, and Ptgds knockdown groups was then assessed, revealing a ~19% increase in the viability of neomycin-treated cells in which Ptgds had been knocked down as compared to those in which it had not (76.9% vs. 57.71%).” This sentence needs to incorporate corresponding figures to describe the results.

• In the second paragraph of section 3.2:

Reviewer #3: Chen and colleagues studied the expression and function of Ptgds in response to neomycin-induced injury. They showed that Ptgds is upregulated in neomycin-injured hair cells and suggested that such upregulation may contribute to the incidence of vertigo and other symptoms of vestibular dysfunction. The authors also found that knocking down Ptgds protects utricle explants and HEI-OC1 cells from neomycin-mediated injury by reducing the production of reactive oxygen species (ROS), the suppression of inflammation, and the abrogation of hair cell apoptosis.

This manuscript tried to highlight the role of Ptgds in aminoglycoside-induced vestibular dysfunction. However, my enthusiasm for this manuscript was significantly diminished by the lack of evidence demonstrating the authors’ proposed mechanism. Please see the attached comment for details.

Reviewer #4: This study aims to investigate the expression and function of Ptgds in HEI-OC1 cells and utricular hair cells, as well as its mechanism of action in aminoglycoside-induced vestibular hair cell damage. The research suggests that Ptgds may be a new target for the prevention and treatment of vestibular dysfunction caused by aminoglycoside drugs. Inhibiting the expression of Ptgds can reduce neomycin-induced apoptosis of vestibular hair cells and enhance hair cell protection. Considering the innovation and clinical significance of the research, this manuscript needs a minor revision before publication.

Minor concerns are as follows:

1.Please ensure that the text format of all images remains consistent. Kindly go through the whole manuscript and make any necessary modification.

2. The damaging concentration of 30mM neomycin may be excessive. Why was 30mM chosen as the optimal damaging concentration in HEI-OC1 cells?

3. How long does neomycin damage the utricle implant? Please supplement it in the text.

4.Please verify if "Ptgds" is spelled correctly. Please go through the whole manuscript and make necessary corrections.

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6. PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy .

Reviewer #1: No

Reviewer #2: Yes:  Yusu Ni

Reviewer #3: No

Reviewer #4: No

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While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

Attachments

Attachment

Submitted filename: Chen_et_al_PLOS ONE_20240521_Shiyi.docx

PONE-D-24-17432R1Ptgds downregulation protect Vestibular hair cells from aminoglycoside-induced vestibulotoxicityPLOS ONE

Dear Dr. Nie,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

ACADEMIC EDITOR:

Following a reviewer, the manuscript still need to clarify the author's hypothesis.

The reviewer could not accept your revision until you make it clear. Please submit your revised manuscript by Sep 20 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Sujeong Jang

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

Reviewer #5: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #5: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #5: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #5: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #5: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: I don't agree with your response for comment 1. Since ototoxic antibiotics could be replaced by many other antibiotics without ototoxic, is this research meaningful? The author should clarify this.

Reviewer #2: The authors have adequately addressed all comments raised, and the manuscript technically sound, and the data support the conclusions, so, I think this manuscript should be accepted for publication.

Reviewer #3: (No Response)

Reviewer #5: (No Response)

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy .

Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

Reviewer #5: Yes:  Byung Chul Kang

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

PONE-D-24-17432R2Ptgds downregulation protect Vestibular hair cells from aminoglycoside-induced vestibulotoxicityPLOS ONE

Dear Dr. Nie,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

==============================

Dear, NieWe have still need to revise the manuscript.As you can see the below, one of our reviewer deeply considered that your manuscript did not complete.Please ensure to revise a reviewer's comment.Thanks.

==============================

Please submit your revised manuscript Dec 20 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Sujeong Jang

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #6: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #6: No

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

Reviewer #6: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #6: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #6: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2:  The authors have adequately addressed all comments raised in a previous round of review and I feel that this manuscript should be acceptable for publication

Reviewer #6:  Overall, this is an interesting review of an important area. However, several areas need attention and I do have a few issues to be addressed by the authors.

1. The meaning of the group was not indicated in figure legend of figure 1C.

2. The conclusion that the expression of Ptgds is upregulated in neomycin-associated injury seems not rigorous bescuse the authors did not verify the result of RNA.

3. The quality of the picture is not high. For example, in figure 3A, the merge of tunel and dapi should be overlapped.

4. Carefully check and improve the English writing in the manuscript.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy .

Reviewer #2: No

Reviewer #6: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

PONE-D-24-17432R3Ptgds downregulation protect Vestibular hair cells from aminoglycoside-induced vestibulotoxicityPLOS ONE

Dear Dr. Nie,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised by the Academic Editor, prior to final decision.

Please submit your revised manuscript by Feb 27 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Miriam A. Hickey, PhD

Academic Editor

PLOS ONE

Editor Comments:

Prior to final decision, please now address these comments from the Academic Editor.

Table 1

Monitor spelling

Please have all genes formatted similarly - the convention being italics for non-human genes.

Please consult here: https://journals.plos.org/plosone/s/submission-guidelines

Use correct and established nomenclature wherever possible.

For all imaging, please provide resolution i.e., actual size of each pixel, with size of z-step, and objective (cellular imaging, immunofluorescence sections).

Immunofluorescence section, please provide RRID number for antibody, if possible.

Please provide numerator, denominator and F values for all ANOVAs (Figure 1B, C, Figure 2 B, C D, Figure 3B, Figure 4 B, C, D and E).

Figure 1C: Please clarify whether cells with Ptgds knockdown were also treated with neo - this is not apparent in figure legend, but text suggests this is the case.

Please provide materials and methods for CCK8 method quoted in figure legends.

What are HEI-Oc1 cells (no information is apparent in materials and methods).

Please clarify/explain the reason for the much lower concentration of neo used for TUNEL experiments (3mM appears to have been used: "samples were treated for 12 h with neomycin (3 mM).").

Figure 3C: This must be analysed with at least 2-way ANOVA as there are two factors (treatments and gene). Please provide numerator, denominator and F value.

Discussion: Please discuss the relevance of the concentrations of neomycin used relative to concentrations observed in clinical samples.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

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Reviewer #2: Yes

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Reviewer #2: Yes

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Reviewer #2: The authors have adequately addressed your comments raised in a previous round of review. We feel that this manuscript should been now acceptable for publication.

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Reviewer #2: No

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PONE-D-24-17432R4Ptgds downregulation protect Vestibular hair cells from aminoglycoside-induced vestibulotoxicityPLOS ONE

Dear Dr. Nie,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Unfortunately, the statistics (F values, numerator, denominator) are not apparent within the submitted supplementary file, which appears to contain Table S1 and Supplementary Figure 1, only.

Therefore, we invite you to submit a revised version of the manuscript that addresses this point.

Please submit your revised manuscript by Mar 29 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to the points above, from the academic editor. You should upload this letter as a separate file labeled 'Response to Editor'.
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We look forward to receiving your revised manuscript.

Kind regards,

Miriam Ann Hickey, PhD

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

Ptgds downregulation protect Vestibular hair cells from aminoglycoside-induced vestibulotoxicity

PONE-D-24-17432R5

Dear Dr. Nie,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Miriam Ann Hickey, PhD

Academic Editor

PLOS ONE

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