PONE-D-22-34859Cytosolic EpCAM cooperates with H-Ras to regulate epithelial to mesenchymal transition through ZEB1PLOS ONE

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Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: No

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

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Reviewer #1: A western blot analysis of EpCAM is suggested to implement figure 1

Indicate the time of experiments shown in figure 2.

Statistical analysis is lacking in all the panel figures.

Proliferation assays at different time points is suggested and a different representation (curve instead of histograms)

Add a western blot analysis of all EMT markers tested in all experiments.

To Assess the efficacy of inhibitors on several signalling pathways a western blot analysis should be included.

Reviewer #2: In this manuscript, the authors investigated the role of cytosolic EpCAM in MCF-10A cell line modified to overexpress the H-Ras oncogene. The manuscript is well written and well structured, with several experiments aimed at defining the interconnected role of H-Ras overexpression and EpCAM in EMT. Figures and graphs are well defined.

There are only a few comments to further improve the work and make it even more complete.

Point 1: The role and possible collateral effects that targeted inhibition of EpCAM would have in epithelial tumors should be expanded.

Point 2: Did you analyze and define the type of cell death involved in the treatment with Paclitaxel and Quercetin? If yes, it would be interesting to add it in the results.

Point 3: In the materials and methods section please add more information about the inhibitors used to test the EpCAM-transformed cells.

Point 4: In the materials and methods section there are some subsections in which the number of cells seeded to perform the experiments has not been included. Please add them.

Point 5: Again, in the materials and methods section, expand the statistical analysis part.

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Reviewer #1: No

Reviewer #2: No

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Submitted filename: PLOSONE PONE-D-22-34859 revision.docx

Cytosolic EpCAM cooperates with H-Ras to regulate epithelial to mesenchymal transition through ZEB1

PONE-D-22-34859R1

Dear Dr. Sankpal,

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.

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Kind regards,

Erika Di Zazzo

Academic Editor

PLOS ONE