FAK downregulation suppresses stem-like properties and migration of human colorectal cancer cells

Chunyan Xu; Wenlu Zhang; Chengxia Liu

doi:10.1371/journal.pone.0284871

Peer Review History

Original SubmissionSeptember 20, 2022
19 Jan 2023 Decision Letter - Salvatore V Pizzo, Editor PONE-D-22-25987FAK downregulation suppresses stem-like properties and migration of human colorectal cancer cellsPLOS ONE Dear Dr. Xu, 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. As you can see both reviewers have raised substantial issues which need to be addressed if the authors plan to resubmit this manuscript. Please submit your revised manuscript by Mar 05 2023 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. 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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: 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: Yes 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: No Reviewer #2: No ****** 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: Dear editor, The manuscript entitled ‘FAK downregulation suppresses stem-like properties and migration of human colorectal cancer cells’ is interesting and novel in the area of cancer. CSCs and their contribution to therapy resistance and tumor progression is without a doubt of value, and the area still needs investigations. The whole manuscript requires edition for grammar and writing errors in order to be a better draft for readers. The sentence ‘cells protein was expression by western blotting analysis’ in the abstract section is wrong. Please rewrite the Abstract in order to be more direct and persuasive. Sentences like ‘Cell viability was measured by cell counting kit-8 and cells protein was expression by western blotting analysis and immunocytochemistry’, ‘Co-expression of FAK and AKT in colorectal cancer cells was assessed by co-immunoprecipitation’, and ‘Spheroid forming ability was evaluated and cell migration measured by wound healing assays’ must not be included in the abstract section. Instead, please refer to the key findings of the study. Introduction. The sentence ‘Both the EMT and the stem cell-like properties of tumor cells promote metastasis’ may pursue a wrong interpretation, as it has known EMT is a feature of cancer stemness not a separate event (10.1007/s12032-022-01801-0). AKT is primarily induced in CSCs through upregulation of the upstream Nanog (J. Clin. Investig. 122 (11) (2012) 4077–4093) or PI3K (Proc. Natl. Acad. Sci. USA 110 (17) (2013) 6829–6834). Where is the place here for FAK? Introduction. Please indicate here that CSCs are equipped with highly efficient DNA repair and effective redox tolerance systems in order to promote therapy resistance (10.1016/j.biopha.2022.113906), and their number is increased outstandingly in cancers with cold immunity (10.1002/jbt.22708). Sincerely Reviewer #2: The present study describes FAK regulation of stem cell-like properties of colon cancer cells. FAK downregulation attenuates such stemness properties and cancer cell migration via AKT phosphorylation. FAK has been target of tumor-suppressor and downregulation of stem cells. The FAK science has well been documented to date. FAK in cytoplasmic region is present in cancer cells and p-AKT is upregulated in EMT, hepatoma and colon cancer metastasis. FAK/AKT/PI3K in EMT and stemness in cancer cells is also well known. The present works are not justified for generalization, indicating a premature submission. 1. More mechanistic explanation in FAK interaction with its partner molecules should be gel-shifted using immuno-precipitation. 2. Several colorectal cancer cells have been analyzed in Fig.1 and etc. The genetic differences in p53 gene are characteristic and the FACK regulated is controlled by p53 and related genes. Therefore, the phenotype properties should be separately examined. 3. PI3K/PTEn coregulation is missed The present study is not acceptable in its present form ****** 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: Cheorl-Ho Kim ******** [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. https://doi.org/10.1371/journal.pone.0284871.r001
Revision 1
30 Jan 2023 Author Response Response to Reviewer #1: Dear reviewer, thank you for your suggestion. 1.The grammar and writing errors of the manuscript have been revised and obtained a language certificate from EditSprings. 2.We have rewritten the Abstract. I hope it could meet your requirement. 3.We have rewritten the sentence "During the initiation of metastasis, epithelial cancer cells undergo an epithelial-mesenchymal transition (EMT), and as a result become migratory and invasive mesenchymal-like cells while acquiring stem cell-like properties and therapy resistance. It was known EMT is a feature of stem cell-like properties in tumor"in Introduction. 4.The AKT/Nanog pathway is critical for maintenance of sarcoma cancer stem cells and spheroid formation. Activation of AKT/β-catenin pathway maintains the growth of cancer stem cells and directly modulates the expression of Nanog and OCT4 in colorectal cancer (doi: 10.1038/s41389-020-00300-z.; doi:10.3892/ijmm.2021.4884). Inconsistently, some studies suggest that AKT is primarily induced in cancer stem cells through upregulation of the upstream Nanog (J. Clin. Investig. 122 (11) (2012) 4077–4093). A cross-regulation Maybe between AKT and Nanog, plays a significant role in maintains stem cell-like properties in tumor. FAK inhibition can suppress ovarian cancer cells migration and invasion through inhibiting downstream PI3K/AKT signaling. (doi: 10.1002/cbin.10184.) 5.We have added the sentence "Cancer stem cells are equipped with highly efficient DNA repair and effective redox tolerance systems in order to promote therapy resistance, and their number is increased outstandingly in cancers with cold immunity" in Introduction. Response to Reviewer #2: Dear reviewer, thank you for your suggestion. 1.In our research, FAK and AKT protein was shown to interact indeed verified by co-immunoprecipitation. 2.In Fig 1, we show the capacity of spheroid formation, cell migration and expression of FAK, p-FAK, AKT, p-AKT, CD44, CD133, OCT4, Nanog and c-Myc in HT29, HCT116, SW480, SW620 and RKO colorectal cancer cells. Expression of p-FAK and p-AKT was higher in HT29 and RKO. In consideration of FAK or AKT was manipulated by transfection experiments and specific inhibitor in subsequent experiments, HT29 and RKO cells were adopted. The results shown in Fig.1 could explain why HT29 and RKO were selected for the subsequent experiments, so the phenotype properties may be dispensable for separately examination. 3.Some studies have reported that the FAK was involved in the regulation of cell mobility via activation of PI3K /AKT pathway, and associated phosphorylation of p85 subunits of tyrosine of PI3K in human cancer cells. (doi:10.1038/nrc3792) ( doi:10.18632/oncotarget.6399). Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0284871.r002
21 Mar 2023 Decision Letter - Salvatore V Pizzo, Editor PONE-D-22-25987R1FAK downregulation suppresses stem-like properties and migration of human colorectal cancer cellsPLOS ONE Dear Dr. Xu, 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. A number of issues were raised by one of the reviewers which should be considered if the authors plan to submit a revised manuscript. Please submit your revised manuscript by May 05 2023 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, Salvatore V Pizzo 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: All comments have been addressed Reviewer #3: 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 #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: 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 #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: 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: The manuscript is acceptable in the current form. All the required amendments are addressed precisely Reviewer #3: - Please carefully check the English grammar, which has many errors and typos. - In Real time-PCR method, what is the thermal cycling conditions? How to calculate and analyze the level of gene expression? These should be specified in the method. - In the results section, the authors have to explain the detail of each result more. - All images of all figures have not the scale bar, it needs to show in the image and also specify in the figure legends. - The authors need to link and interpret the data in the results and discussion about the two different cell lines, HT29 and RKO. ****** 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 #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. https://doi.org/10.1371/journal.pone.0284871.r003
Revision 2
28 Mar 2023 Author Response Dear reviewers, Thank you for your letter and for the reviewer’ comments concerning our manuscript. Those comments are all valuable and very helpful for revising and improving our paper. We have studied comments carefully and have made correction which we hope meet with approval. Reviewer #3: 1. Please carefully check the English grammar, which has many errors and typos. We invited the high qualified and native English speakers at EditSprings again to modify the English grammar in our manuscript. 2. In Real time-PCR method, what is the thermal cycling conditions? How to calculate and analyze the level of gene expression? These should be specified in the method. We added the thermal cycling conditions and the formula to calculate and analyze the level of gene expression in the method section. 3. In the results section, the authors have to explain the detail of each result more. We made some changes in the results section. We hope it could meet your requirement. 4. All images of all figures have not the scale bar, it needs to show in the image and also specify in the figure legends. We added the scale bar in all images of all figures. I hope it could meet your requirement. 5. The authors need to link and interpret the data in the results and discussion about the two different cell lines, HT29 and RKO. In the discussion section, we have linked and interpreted the data of the results. HT29 and RKO are all colorectal cancer cell lines, so we did not discuss them separately. We sincerely hope that this revised manuscript has addressed all your comments and suggestions. We appreciated for reviewer’ warm work earnestly, and hope that the correction will meet with approval. Once again, thank you very much for your comments and suggestions. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0284871.r004
5 Apr 2023 Decision Letter - Salvatore V Pizzo, Editor PONE-D-22-25987R2FAK downregulation suppresses stem-like properties and migration of human colorectal cancer cellsPLOS ONE Dear Dr. Xu, 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. Your manuscript is basically acceptable for publication but one reviewer has indicated a labelling error. It is preferable for the authors to correct this if the reviewer is correct rather than editorial change of the legend. Please submit your revised manuscript by May 20 2023 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, Salvatore V Pizzo 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. [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 #3: 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 #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? 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 #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 #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 #3: All images of all figures have already added the scale bar, but in the figure legends are not correct. Authors only wrote the word "(magnification, ×40)", so it should be written "(magnification, ×40; scale bar 20 μm)". Please edit and rewrite them all of the figure legends. ****** 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 #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. https://doi.org/10.1371/journal.pone.0284871.r005
Revision 3
5 Apr 2023 Author Response Dear reviewer, Thank you for your letter and for the reviewer’ comments concerning our manuscript. This comment is valuable and very helpful for revising and improving our paper. We have studied the comment carefully and have made correction which we hope meet with approval. Reviewer #3: All images of all figures have already added the scale bar, but in the figure legends are not correct. Authors only wrote the word "(magnification, ×40)", so it should be written "(magnification, ×40; scale bar 20 μm)". Please edit and rewrite them all of the figure legends. We edit and rewrite the figure legends as your suggestion. I hope it could meet your requirement. We sincerely hope that this revised manuscript has addressed all your comments and suggestions. We appreciated for reviewer’ warm work earnestly, and hope that the correction will meet with approval. Once again, thank you very much for your comments and suggestions. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0284871.r006
11 Apr 2023 Decision Letter - Salvatore V Pizzo, Editor FAK downregulation suppresses stem-like properties and migration of human colorectal cancer cells PONE-D-22-25987R3 Dear Dr. Xu, 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. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. 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, Salvatore V Pizzo Academic Editor PLOS ONE Additional Editor Comments (optional): 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 #3: 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 #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? 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 #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 #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 #3: (No Response) ****** 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 #3: No ******** https://doi.org/10.1371/journal.pone.0284871.r007
Formally Accepted
13 Apr 2023 Acceptance Letter - Salvatore V Pizzo, Editor PONE-D-22-25987R3 FAK downregulation suppresses stem-like properties and migration of human colorectal cancer cells Dear Dr. Xu: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. 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. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Salvatore V Pizzo Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0284871.r008

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