Dear Dr. Petyim,

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.

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We look forward to receiving your revised manuscript.

Kind regards,

Stefania Crispi

Academic Editor

PLOS ONE

Journal requirements:

When submitting your revision, we need you to address these additional requirements.

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

2. In the online submission form, you indicated that all the data analyzed during this study are available from the corresponding author upon reasonable request.

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3. Thank you for stating the following financial disclosure:

This study was supported by the Research Development Grants, Faculty of

Medicine Siriraj Hospital, Mahidol University (no. R016631035).

Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."

If this statement is not correct you must amend it as needed.

Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf.

4. Please include a separate caption for each figure in your manuscript.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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

Reviewer #1: Yes

Reviewer #2: Partly

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

Reviewer #1: Yes

Reviewer #2: Yes

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

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: The authors with improved methodological transparency, statistical justification, and language editing, the manuscript could make a strong contribution to the field. The biological insights are interesting and potentially impactful, but the presentation and interpretation need refinement for clarity.

The introduction effectively summarizes the background but repeats information (e.g., progestin effects on apoptosis). Please streamline to highlight the knowledge gap specifically, why drospirenone’s distinct chemical structure warrants this study.

The “Materials and Methods” section provides substantial procedural detail, but lacks clarity on certain aspects:

- Specify the number of biological replicates and technical replicates for each assay.

- Clarify the rationale for selecting 1 µM drospirenone—mention how this concentration relates to physiological or pharmacological levels.

- Indicate how cell purity was verified (e.g., immunostaining for stromal markers such as vimentin or absence of epithelial markers).

- Has a characterization of the cells isolated from the patients been done?

While statistical methods are mentioned, there is no justification for using parametric tests with such small sample sizes (n=8 effective samples). Consider verifying data normality or using non-parametric tests. Include sample size and variance information in figure legends.

Figures and results are clearly structured, but key data need better quantification:

- Report exact p-values instead of only “p < 0.05”.

- Include mean ± SEM or box plots for all bar charts.

- Describe Figure 6 results more precisely—state whether fold changes were normalized to controls.

The discussion is comprehensive but overly descriptive and partially redundant.

- Clearly differentiate between your findings and those from previous studies.

- The explanation of apoptotic pathways (intrinsic vs extrinsic) is useful but needs more direct connection to your gene-expression data.

- Include a short paragraph on clinical relevance (e.g., how these in vitro results may inform treatment regimens).

The manuscript is generally understandable but requires language polishing to meet journal standards.

- Remove unnecessary periods after abbreviations (e.g., “.In many in vitro studies” → “In many in vitro studies”).

- Correct spacing and grammar inconsistencies (“10%.DMEM” → “10% DMEM”).

- Consider professional English editing for smoother readability.

Minor Comments

1. Line 70–74: “Endometriosis is a complex gynecological condition...” → Combine short sentences for better flow.

2. Line 105–120: Clarify inclusion/exclusion criteria and ethical consent procedures.

3. Table 1: The forward primer for BCL2 is missing; check for duplication (“R:” repeated twice).

4. Figures: Ensure consistent file naming (“Figure 6” not “Figure” only).

5. Add abbreviation list after the abstract or before references (e.g., EU-ESCs, EC-ESCs, DRSP).

6. Reference section: Check formatting according to PLOS ONE guidelines journal names should be italicized and consistent.

Reviewer #2: The article is interetsing and presents original data. I have some concerns that should be addressed by the authors; the manuscript should be modified and implemented accordingly

1) The authors should perform some preliminary studies on the isolated cells in order to better define their phenotype, such as analyze the expression of estrogen and progesteron, and expression of stromal markers such as CD10. Moreover, to investigate the effect of immortalization on cell karyotypes, cytogenetic analysis should be performed using standard

G-banding analysis

2) In order to better define the effect of Drospirenone, I suggest to define effects on cell cycle by FACS and the effects on celle mobility by a migration assay

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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

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To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures

You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation.

NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications.

<p>Drospirenone promotes apoptosis in ectopic but inhibits proliferation in eutopic human endometrial stromal cells

PONE-D-25-51558R1

Dear Dr. Somsin Petyim,

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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If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Stefania Crispi

Academic Editor

PLOS One

Additional Editor Comments (optional):

All the reviewers comments have been addressed

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

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

Reviewer #1: (No Response)

Reviewer #2: Yes

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

Reviewer #1: (No Response)

Reviewer #2: I Don't Know

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

The PLOS Data policy

Reviewer #1: (No Response)

Reviewer #2: Yes

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

Reviewer #1: (No Response)

Reviewer #2: Yes

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Reviewer #1: (No Response)

Reviewer #2: (No Response)

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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: Alfonso Baldi

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