PONE-D-24-13072Monocarboxylate transporter dependent mechanism is involved in proliferation, migration, and invasion of human glioblastoma cell lines via activation of PI3K/Akt signalingpathwayPLOS ONE

Dear Dr. Gao,

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.

Reviewer 1: Major points - The scientific background in the introduction section is very poor. More information is needed to give a clear overview of the current state of knowledge and the novelty of the results reported in the manuscript. - How do you explain the very small increase of MCT4 in the “overexpressing cells”, considering that it is almost the same in all control cell lines (blank(?), control siRNA, empty vector)? - Based on the biochemical role of the MCT4 transporter, the correct characterization of the cell line models generated here requires the metabolic profiles of these cells (e.g. intracellular pH, lactate levels, ECAR/OCR levels, ATP production). - Lines 48-50: please, clarify this statement. - Considering that MCT4 regulates the transport of lactate across the membrane and modulates the mitochondrial activity of cells, the MTT assay is completely inappropriate to test cell viability. To detect the effects of MCT4 expression on cell proliferation and cell cycle distribution, more appropriate assays such as the trypan blue assay, cytofluorimetry and the BrdU/EdU assay are required. - Based on the previous findings discussed in the manuscript, measurement of VEGF production may be useful. - To demonstrate the effect of the MCT4/CD147 axis on the PI3K/Akt signaling pathway, a simple analysis of Akt phosphorylation is not sufficient. Based on previous findings (Zhou Y, 2020; Bu X, 2021), the authors should improve the results to substantiate their findings. The levels of phospho-GSK3b/GSK3b and Nrf2 should be checked. - Considering that both Akt and ATM/ATR inhibitors are available (and there are no selective MCT4 inhibitors to date, Goldberg WF, 2023), assessing the differential sensitivity of the described cancer cell models to these agents could be of value for a translational impact of the results presented here. - The reference list is insufficient. Minor points - Line 180: What do you mean by “no drug treatment”? - Blank: is an invalid control definition, please replace it in the text, legend and figures - Results of relative quantitative analysis by immunoblotting: please explain better how you performed it - Figure 4: Quantification of Akt phosphorylation should be expressed as the ratio of P-Akt band density/Akt band density relative to actin level. - Methods: There is not enough information to reproduce some of the experiments. Please provide more details for Western blot and immunofluorescence analysis. Add a detailed description of the invasion assay where the removal of the Matrigel is not mentioned. - Paragraph 3.3: Explain better the results of migration and invasion. Give the values as a percentage reduction of the wound compared to the corresponding control. - Remove the comments from all figure legends and explain the technical details of the experiments shown (what they represent and how the values given in the graphs were calculated). - Please provide the originals of the immunoblot analyses. 

Reviewer 2: In this manuscript authors clarify the role of the pH regulator monocarboxylate transporter MCT4 and its accessory h-subunit CD147 in glioblastoma cells. Briefly, they demonstrated that inhibiting the activity and expression of the MCT4/CD147 transporter complex decreases cell proliferation, migration, invasion, and Akt activation, while lentivirus-mediated MCT4 overexpression reverts the system in glioblastoma cells. Manuscript needs some minor and major revisions: - An extensive English editing is required. I suggest contacting a native speaker. - There are several typing errors in the whole text. Please, check them in the whole text. - Genes must be written in italic. - The acronyms in the abstract should be replaced with the relative extended. - Introduction is too short and assume several concepts that could be unknown for the readers. To this, I suggest improving the background section. - Densitometry of MCT4(43KD)U251 in “Control siRNA” sample (Fig.1B) appears not to correspond to the band intensity of the relative WB (Fig.1A). Please check it. - The quality of Western Blots analyses is poor. All Wb are oversaturated. Please replace them with blots with reduced exposure, when it’s possible, and add the original blots as supplementary materials. - To test the invasion ability of glioblastoma cells, authors have performed the transwell assay. However, the 3D spheroid invasion assay may represent the most relevant assay for this goal, particularly for glioblastoma cell lines, as well as to better mimic tumor behavior in vivo [doi: 10.3791/52686]. Briefly, 3D tumor spheroids are embedded into 3D ECM. It is expected to see that non-invasive cancer cells stay as compact spheroids with a distinct border to the surrounding ECM and do not show any obvious signs of invasion. On the contrary, invasive cells start to invade into the surrounding matrix and display outgrowth from the spheroids [doi: 10.1371/journal.pone.0293475; doi: 10.3389/fcell.2023.1272667]. - What is the difference between glycosylated and non-glycosylated CD147 (Fig. 2A)? Please explain this concept in more depth. - In the Discussion section, authors assume the involvement of PI3K/Akt signaling pathway. However, they have performed only the WB analysis of Akt protein expression without verify the involvement of PI3K or other key players in this pathway. I highly recommend exploring the other components or activation of downstream molecules.

Please submit your revised manuscript by Aug 07 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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We look forward to receiving your revised manuscript.

Kind regards,

Nicola Amodio, PhD

Academic Editor

PLOS ONE

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Additional Editor Comments:

Dear Dr. Gao,

two reviewers have assessed your manuscript.

Although findings look interesting, both reviewers are asking major revisions to enhance the quality of the manuscript.

Reviewer 1:

Major points

- The scientific background in the introduction section is very poor. More information is needed to give a clear overview of the current state of knowledge and the novelty of the results reported in the manuscript.

- How do you explain the very small increase of MCT4 in the “overexpressing cells”, considering that it is almost the same in all control cell lines (blank(?), control siRNA, empty vector)?

- Based on the biochemical role of the MCT4 transporter, the correct characterization of the cell line models generated here requires the metabolic profiles of these cells (e.g. intracellular pH, lactate levels, ECAR/OCR levels, ATP production).

- Lines 48-50: please, clarify this statement.

- Considering that MCT4 regulates the transport of lactate across the membrane and modulates the mitochondrial activity of cells, the MTT assay is completely inappropriate to test cell viability. To detect the effects of MCT4 expression on cell proliferation and cell cycle distribution, more appropriate assays such as the trypan blue assay, cytofluorimetry and the BrdU/EdU assay are required.

- Based on the previous findings discussed in the manuscript, measurement of VEGF production may be useful.

- To demonstrate the effect of the MCT4/CD147 axis on the PI3K/Akt signaling pathway, a simple analysis of Akt phosphorylation is not sufficient. Based on previous findings (Zhou Y, 2020; Bu X, 2021), the authors should improve the results to substantiate their findings. The levels of phospho-GSK3b/GSK3b and Nrf2 should be checked.

- Considering that both Akt and ATM/ATR inhibitors are available (and there are no selective MCT4 inhibitors to date, Goldberg WF, 2023), assessing the differential sensitivity of the described cancer cell models to these agents could be of value for a translational impact of the results presented here.

- The reference list is insufficient.

Minor points

- Line 180: What do you mean by “no drug treatment”?

- Blank: is an invalid control definition, please replace it in the text, legend and figures

- Results of relative quantitative analysis by immunoblotting: please explain better how you performed it

- Figure 4: Quantification of Akt phosphorylation should be expressed as the ratio of P-Akt band density/Akt band density relative to actin level.

- Methods: There is not enough information to reproduce some of the experiments. Please provide more details for Western blot and immunofluorescence analysis. Add a detailed description of the invasion assay where the removal of the Matrigel is not mentioned.

- Paragraph 3.3: Explain better the results of migration and invasion. Give the values as a percentage reduction of the wound compared to the corresponding control.

- Remove the comments from all figure legends and explain the technical details of the experiments shown (what they represent and how the values given in the graphs were calculated).

- Please provide the originals of the immunoblot analyses.

Reviewer 2:

In this manuscript authors clarify the role of the pH regulator monocarboxylate transporter MCT4 and its accessory h-subunit CD147 in glioblastoma cells. Briefly, they demonstrated that inhibiting the activity and expression of the MCT4/CD147 transporter complex decreases cell proliferation, migration, invasion, and Akt activation, while lentivirus-mediated MCT4 overexpression reverts the system in glioblastoma cells.

Manuscript needs some minor and major revisions:

- An extensive English editing is required. I suggest contacting a native speaker.

- There are several typing errors in the whole text. Please, check them in the whole text.

- Genes must be written in italic.

- The acronyms in the abstract should be replaced with the relative extended.

- Introduction is too short and assume several concepts that could be unknown for the readers. To this, I suggest improving the background section.

- Densitometry of MCT4(43KD)U251 in “Control siRNA” sample (Fig.1B) appears not to correspond to the band intensity of the relative WB (Fig.1A). Please check it.

- The quality of Western Blots analyses is poor. All Wb are oversaturated. Please replace them with blots with reduced exposure, when it’s possible, and add the original blots as supplementary materials.

- To test the invasion ability of glioblastoma cells, authors have performed the transwell assay. However, the 3D spheroid invasion assay may represent the most relevant assay for this goal, particularly for glioblastoma cell lines, as well as to better mimic tumor behavior in vivo [doi: 10.3791/52686]. Briefly, 3D tumor spheroids are embedded into 3D ECM. It is expected to see that non-invasive cancer cells stay as compact spheroids with a distinct border to the surrounding ECM and do not show any obvious signs of invasion. On the contrary, invasive cells start to invade into the surrounding matrix and display outgrowth from the spheroids [doi: 10.1371/journal.pone.0293475; doi: 10.3389/fcell.2023.1272667].

- What is the difference between glycosylated and non-glycosylated CD147 (Fig. 2A)? Please explain this concept in more depth.

- In the Discussion section, authors assume the involvement of PI3K/Akt signaling pathway. However, they have performed only the WB analysis of Akt protein expression without verify the involvement of PI3K or other key players in this pathway. I highly recommend exploring the other components or activation of downstream molecules.

[Note: HTML markup is below. Please do not edit.]

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: No

Reviewer #2: Partly

**********

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

Reviewer #1: Yes

Reviewer #2: 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: No

Reviewer #2: Yes

**********

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: Yes

Reviewer #2: Yes

**********

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: In this manuscript, the authors describe the role of the MCT4 transporter in glioblastoma. They hypothesize that the MCT4/CD147 complex promotes proliferation, migration and invasion by activating the Akt signaling pathway. The rationale for the experiments performed is quite good, but in my opinion some points should be addressed to improve the quality of the manuscript. The author's statement in the discussion in lines 344-351 can hardly be supported by the manuscript in its present form.

Major points

- The scientific background in the introduction section is very poor. More information is needed to give a clear overview of the current state of knowledge and the novelty of the results reported in the manuscript.

- How do you explain the very small increase of MCT4 in the “overexpressing cells”, considering that it is almost the same in all control cell lines (blank(?), control siRNA, empty vector)?

- Based on the biochemical role of the MCT4 transporter, the correct characterization of the cell line models generated here requires the metabolic profiles of these cells (e.g. intracellular pH, lactate levels, ECAR/OCR levels, ATP production).

- Lines 48-50: please, clarify this statement.

- Considering that MCT4 regulates the transport of lactate across the membrane and modulates the mitochondrial activity of cells, the MTT assay is completely inappropriate to test cell viability. To detect the effects of MCT4 expression on cell proliferation and cell cycle distribution, more appropriate assays such as the trypan blue assay, cytofluorimetry and the BrdU/EdU assay are required.

- Based on the previous findings discussed in the manuscript, measurement of VEGF production may be useful.

- To demonstrate the effect of the MCT4/CD147 axis on the PI3K/Akt signaling pathway, a simple analysis of Akt phosphorylation is not sufficient. Based on previous findings (Zhou Y, 2020; Bu X, 2021), the authors should improve the results to substantiate their findings. The levels of phospho-GSK3b/GSK3b and Nrf2 should be checked.

- Considering that both Akt and ATM/ATR inhibitors are available (and there are no selective MCT4 inhibitors to date, Goldberg WF, 2023), assessing the differential sensitivity of the described cancer cell models to these agents could be of value for a translational impact of the results presented here.

- The reference list is insufficient.

Minor points

- Line 180: What do you mean by “no drug treatment”?

- Blank: is an invalid control definition, please replace it in the text, legend and figures

- Results of relative quantitative analysis by immunoblotting: please explain better how you performed it

- Figure 4: Quantification of Akt phosphorylation should be expressed as the ratio of P-Akt band density/Akt band density relative to actin level.

- Methods: There is not enough information to reproduce some of the experiments. Please provide more details for Western blot and immunofluorescence analysis. Add a detailed description of the invasion assay where the removal of the Matrigel is not mentioned.

- Paragraph 3.3: Explain better the results of migration and invasion. Give the values as a percentage reduction of the wound compared to the corresponding control.

- Remove the comments from all figure legends and explain the technical details of the experiments shown (what they represent and how the values given in the graphs were calculated).

- Please provide the originals of the immunoblot analyses.

Reviewer #2: In this manuscript authors clarify the role of the pH regulator monocarboxylate transporter MCT4 and its accessory h-subunit CD147 in glioblastoma cells. Briefly, they demonstrated that inhibiting the activity and expression of the MCT4/CD147 transporter complex decreases cell proliferation, migration, invasion, and Akt activation, while lentivirus-mediated MCT4 overexpression reverts the system in glioblastoma cells.

Manuscript needs some minor and major revisions:

- An extensive English editing is required. I suggest contacting a native speaker.

- There are several typing errors in the whole text. Please, check them in the whole text.

- Genes must be written in italic.

- The acronyms in the abstract should be replaced with the relative extended.

- Introduction is too short and assume several concepts that could be unknown for the readers. To this, I suggest improving the background section.

- Densitometry of MCT4(43KD)U251 in “Control siRNA” sample (Fig.1B) appears not to correspond to the band intensity of the relative WB (Fig.1A). Please check it.

- The quality of Western Blots analyses is poor. All Wb are oversaturated. Please replace them with blots with reduced exposure, when it’s possible, and add the original blots as supplementary materials.

- To test the invasion ability of glioblastoma cells, authors have performed the transwell assay. However, the 3D spheroid invasion assay may represent the most relevant assay for this goal, particularly for glioblastoma cell lines, as well as to better mimic tumor behavior in vivo [doi: 10.3791/52686]. Briefly, 3D tumor spheroids are embedded into 3D ECM. It is expected to see that non-invasive cancer cells stay as compact spheroids with a distinct border to the surrounding ECM and do not show any obvious signs of invasion. On the contrary, invasive cells start to invade into the surrounding matrix and display outgrowth from the spheroids [doi: 10.1371/journal.pone.0293475; doi: 10.3389/fcell.2023.1272667].

- What is the difference between glycosylated and non-glycosylated CD147 (Fig. 2A)? Please explain this concept in more depth.

- In the Discussion section, authors assume the involvement of PI3K/Akt signaling pathway. However, they have performed only the WB analysis of Akt protein expression without verify the involvement of PI3K or other key players in this pathway. I highly recommend exploring the other components or activation of downstream molecules.

**********

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: 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-24-13072R1Monocarboxylate transporter dependent mechanism is involved in proliferation, migration, and invasion of human glioblastoma cell lines via activation of PI3K/Akt signalingpathwayPLOS ONE

Dear Dr. Gao,

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. - Please define MCT, CD147, PBKVARt, VEGF, and siRNA acronyms at their first mention in both the abstract and the introduction.

- Line 73: Correct "lacate" to "lactate".

- British or American spelling (e.g., “tumor” vs. “tumour”) should be selected and used consistently throughout the manuscript. -Please better address reviewer's 2 questions.

Please submit your revised manuscript by Nov 18 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,

Nicola Amodio, PhD

Academic Editor

PLOS ONE

[Note: HTML markup is below. Please do not edit.]

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: (No Response)

Reviewer #2: All comments have been addressed

Reviewer #3: (No Response)

**********

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: Partly

Reviewer #2: Yes

Reviewer #3: Partly

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: 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

**********

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: No

Reviewer #2: Yes

Reviewer #3: 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 appreciate the authors' efforts to improve their results and the quality of the manuscript. However, they have not addressed most, if not all, of the functional issues raised. In my opinion, the results presented are insufficient to support the findings claimed by the authors.

Reviewer #2: Since authors have performed all the changes requested by the reviewers, the manuscript is now acceptable for publication.

Reviewer #3: In this study. Dr. Gao and colleagues aim to clarify the mechanisms underlying MCT4/CD147 role in Glioblastoma progression and aggressiveness in human glioblastoma in vitro models. The manuscript was already reviewed and modified accordingly but I might suggest something else to revise before publication.

- Please define MCT, CD147, PBKVARt, VEGF, and siRNA acronyms at their first mention in both the abstract and the introduction.

- Line 73: Correct "lacate" to "lactate".

- British or American spelling (e.g., “tumor” vs. “tumour”) should be selected and used consistently throughout the manuscript.

- Please consider including more specific information in the transwell method section about the statistical methods used to validate experimental results. Moreover, the added part after revision seems to be taken directly from a protocol and not contextualized.

- Ensure consistency when referring to “siRNA interference” or “siRNA knockdown.”

- The revised manuscript includes raw Western blot images, as requested. Make sure that figure legends are adequately described for example by adding how images were quantified using ImageJ (mentioned in the Results but could be added in the captions as well).

- Consider adding some recent (last 2 years) literature on MCT4/CD147 role in glioblastoma as references.

- Please also state the rationale for using ANOVA with post hoc tests, and if it is well justified for your specific requirements and data.

- The revised discussion has improved but can still be more cautious in the mechanistic conclusions you made. For example, when discussing MCT/ CD147 and Akt signaling, consider removing the assumption with something like suggestion or hypothesis.

Overall, my suggestion to the editor is Minor Revisions.

**********

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: Yes: Anna Martina Battaglia

Reviewer #3: 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.

Monocarboxylate transporter dependent mechanism is involved in proliferation, migration, and invasion of human glioblastoma cell lines via activation of PI3K/Akt signalingpathway

PONE-D-24-13072R2

Dear Dr. Gao,

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,

Nicola Amodio, PhD

Academic Editor

PLOS ONE

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