Rebuttal Letter

All the significant change was marked in blue.

Responses to Editor-in-Chief’s Comments

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

• Author’s Response: Thank you for your feedback on the style requirements for PLOS ONE. We have updated our manuscript to comply with the necessary templates and apologize for any inconvenience caused. We appreciate your efforts in maintaining high standards in scientific publishing.

2. Thank you for stating the following financial disclosure:

“NO - Include this sentence at the end of your statement: The funders had no role in study design, data collection, and analysis, the decision to publish, or preparation of the manuscript.”

At this time, please address the following queries:

a) Please clarify the sources of funding (financial or material support) for your study. List the grants or organizations that supported your study, including funding received from your institution.

b) State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection, and analysis, the decision to publish, or preparation of the manuscript.”

c) If any authors received a salary from any of your funders, please state which authors and which funders.

d) If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.”

Please include your amended statements within your cover letter; we will change the online submission form on your behalf.

• Author’s Response: Thank you for the suggestion. As authors, we value your input and have considered it. We have carefully revised the financial disclosure of our paper in the revised version and also included the necessary details in the cover letter.

3. In your Data Availability statement, you have not specified where the minimal data set underlying the results described in your manuscript can be found. PLOS defines a study's minimal data set as the underlying data used to reach the conclusions drawn in the manuscript and any additional data required to replicate the reported study findings in their entirety. All PLOS journals require that the minimal data set be made fully available. For more information about our data policy, please see http://journals.plos.org/plosone/s/data-availability.

Upon re-submitting your revised manuscript, please upload your study’s minimal underlying data set as either Supporting Information files or to a stable, public repository and include the relevant URLs, DOIs, or accession numbers within your revised cover letter. For a list of acceptable repositories, please see http://journals.plos.org/plosone/s/data-availability#loc-recommended-repositories. Any potentially identifying patient information must be fully anonymized.

Important: If there are ethical or legal restrictions to sharing your data publicly, please explain these restrictions in detail. Please see our guidelines for more information on what we consider unacceptable restrictions to publicly sharing data: http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions. Note that it is not acceptable for the authors to be the sole named individuals responsible for ensuring data access.

We will update your Data Availability statement to reflect the information you provide in your cover letter.

• Author’s Response: As authors, we appreciate the suggestion for a Data Availability statement. In our revised paper, we have mentioned that all relevant data are included in the manuscript and its Supporting Information files.

4. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide.

• Author’s Response: Thank you for your feedback on our manuscript. We appreciate your suggestion for a Data Availability statement and would like to confirm that we have provided all relevant data in the manuscript and its Supporting Information files. We do not have any additional data to provide. Please note that we have updated the Data Availability statement in our paper submission to reflect this information. We apologize for any confusion caused by our previous statement and hope that this revised statement meets the requirements of your journal.

5. PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6th, 2016. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager. To do this, go to ‘Update my Information’ (in the upper left-hand corner of the main menu), and click on the Fetch/Validate link next to the ORCID field. This will take you to the ORCID site and allow you to create a new ID or authenticate a pre-existing iD in Editorial Manager. Please see the following video for instructions on linking an ORCID iD to your Editorial Manager account: https://www.youtube.com/watch?v=_xcclfuvtxQ

• Author’s Response: Thank you for notifying us of the ORCID iD requirement for the corresponding author in Editorial Manager. We confirm that our corresponding author has an ORCID iD and we ensured that it is validated in Editorial Manager as per your instructions.

6. Please amend the manuscript submission data (via Edit Submission) to include author Junjiang Fu.

• Author’s Response: Thank you for your email. We confirm that we would like to add Junjiang Fu as an author to our manuscript. We amended the manuscript submission data as requested via the Edit Submission option.

We appreciate your attention to detail and apologize for any confusion caused by the omission of Prof. Fu's name. Thank you for your assistance in this matter.

7. Your ethics statement should only appear in the Methods section of your manuscript. If your ethics statement is written in any section besides the Methods, please move it to the Methods section and delete it from any other section. Please ensure that your ethics statement is included in your manuscript, as the ethics statement entered into the online submission form will not be published alongside your manuscript.

• Author’s Response: We confirm that we have included an ethics statement in our manuscript, but it may have been included in a section other than the Methods. We moved the ethics statement to the Methods section and delete it from any other section, as per your instructions. We understand the importance of properly including our ethics statement and ensuring its visibility in the published manuscript. Thank you for bringing this to our attention and for your continued support.

8. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels.

• In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions.

• Author’s Response: We would like to thank the reviewers for their valuable comments on our manuscript. In response to their suggestions, we have thoroughly reviewed our blot and gel images and made necessary adjustments to ensure they comply with the blot/gel reporting and figure preparation requirements outlined by PLOS ONE. We have provided the original uncropped and unadjusted images underlying all blot or gel results reported in Supporting Information files as “S1_raw_images”, as required by the journal. Additionally, we would like to note that all our blot/gel image data are available in the Supporting Information files. We hope that our revised manuscript meets the requirements of PLOS ONE and thank the editor and reviewers for their time and effort in reviewing our work.

9. 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.

• Author’s Response: Thank you for bringing this matter to our attention. We have thoroughly reviewed our reference list and can confirm that there are no references to retracted papers in our manuscript. We appreciate your guidance in this matter and are confident that our reference list is complete and accurate. We ensured that any changes made to the reference list are mentioned in the accompanying rebuttal letter. Thank you once again for your valuable feedback.

Responses to Reviewer's #1 comments

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: Yes

• Author’s Response: The author extends their heartfelt appreciation for the encouraging feedback and unwavering support they have received for their work. They are truly honored and humbled by the positive input and feel privileged to have received such a warm reception. The author values the opportunity to contribute to the field and is grateful for the review, which provides inspiration for their future endeavors.

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

Reviewer #1: Yes

• Author’s Response: Thank you very much, Reviewer #1, for confirming that our statistical analysis has been performed appropriately and rigorously. We have made every effort to ensure that our methodology is sound and that our results are reliable. Your positive feedback is greatly appreciated.

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 exceptions (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 in 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

• Author’s Response: Thank you, Reviewer #1, for confirming that we have made all data underlying the findings in our manuscript fully available. We have made every effort to comply with the PLOS Data policy and have provided our data as part of the manuscript or its supporting information. We have also ensured that participant privacy and any restrictions on sharing data have been clearly specified where applicable. We appreciate your positive feedback on this matter.

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

• Author's Response: Thank you, Reviewer #1, for confirming that our manuscript is presented in an intelligible fashion and written in standard English. We have made every effort to ensure that our language is clear, correct, and unambiguous, and we are grateful for your positive feedback. We continue to carefully proofread and edit our manuscript to correct any typographical or grammatical errors before submission.

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: This paper contains interesting findings that examine the involvement of chemokines in melanoma progression and their mechanism. However, the following points need to be clarified.

• Which isoform is referred to in the experimental results when only "DMKN" is mentioned? This should be clear, as the action may differ from isoform to isoform.

o Author's Response: We thank Reviewer #1 for their insightful feedback and agree that the lack of clarity in our manuscript regarding which DMKN isoform is referred to in the experimental results could have been improved. We appreciate your attention to this matter, and we have considered your comments in our revisions. As we mentioned in Fig. 2A and 2B, the expression of DMKN-β genes was almost completely lost in the DMKN-shRNA-transduced cells (shDMKN) compared to the lentivirus-scrambled control (NC). Accordingly, these cell lines were selected for future in vivo analysis. In our study, we used DMKN-β isoforms, which are the main expressed isoforms of DMKN in metastatic melanoma. To address your concern, we have revised our manuscript to clarify which DMKN isoform is being referred to in each experiment. Specifically, we have added details in the Results section to specify which isoform was used in each experiment. Additionally, we have mentioned in the revised Discussion section that DMKN-β isoforms were used in the study and that we have clarified which isoform is being referred to in each experiment. We hope that these revisions make our manuscript clearer and more informative, and we appreciate your valuable feedback.

• Page 21. The difference between low-DMKN and high-DMKN in the following statement is unclear.

"Given both DMKN mRNA and protein levels, 48.39% (15) of patients were classified as low-DMKN(FC < 0.18 for DMKN mRNA expression and > 42.78 μg/L for serum DMKN levels), while almost 51.61% (16) were grouped as high-DMKN (FC≤0.18 for DMKN mRNA expression and ≥ 42.78 μg/L for protein DMKN levels."

o Author's Response: Thank you, Reviewer, for bringing this to our attention. We apologize for any confusion caused by the unclear difference between low-DMKN and high-DMKN in the statement you highlighted. We revised the manuscript to clarify that low-DMKN patients had an FC (fold change) value less than 0.18 for DMKN mRNA expression and a serum DMKN level greater than 42.78 μg/L, while high-DMKN patients had an FC value less than or equal to 0.18 for DMKN mRNA expression and a serum DMKN level equal to or greater than 42.78 μg/L. We appreciate your feedback and ensure that this is addressed in the revised manuscript.

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