PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cells

Ji On Kim; Seung Eun Baek; Eun Yeong Jeon; Jong Min Choi; Eun Jeong Jang; Chi Dae Kim

doi:10.1371/journal.pone.0265191

Peer Review History

Original SubmissionJuly 4, 2021
1 Dec 2021 Decision Letter - Suresh Kumar Verma, Editor PONE-D-21-21832PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cellsPLOS ONE Dear Dr. Kim, 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. The reviewers express concerns with the experimental design and result quality. the role of PDGFR-b mediate HMGB1 effects on smooth muscle cells should be included. Appropriate references needs to update and method section require more clarity. Please submit your revised manuscript within 30 days. 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, Suresh Kumar Verma, PhD 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. 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 3. 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 4. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section 5. 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 [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 Reviewer #3: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: I Don't Know Reviewer #2: Yes Reviewer #3: 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 Reviewer #3: 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 Reviewer #3: 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 the present study, the authors studied signaling mediating release of HMGB1, a damage-associated molecular which mediates vascular complications in stress vasculature, in mechanically stressed vascular smooth muscle cells (VSMCs) isolated from mouse aorta SD rats. They found that HMGB1 released from stressed VSMCs was increased which accompanied with increased cytosolic translocation of nuclear HMGB1, and acetylated and phosphorylated form of HMGB1. Furthermore, they found that PDGFR inhibitor AG1295 inhibited HMGB1 release in stressed VSMCs. Moreover the increased HMGB1 release from stressed VSMCs was diminished when PDGFR-β was silenced by siRNA. PDGF-DD increased HMGB1 release. Finally the authors demonstrated that PDGFR-β-mediated secretion of HMGB1 in VSMCs might be a promising therapeutic strategy for hypertension-related vascular complications. The major concern of this manuscript is that the authors did not provide any evidence showing that the role of PDGFR-β/p-PDGFR-β-mediated HMGB1 in biology and function (i.e. contractile property) of VSMCs which makes the conclusion very weak. Other concerns including: 1. References should be added regarding the prerequisite of the cytosolic translation of nuclear HMGB1 for the secretion of HMGB1. 2. How p-PDGFR-β regulates HMGB1 cytosolic translocation and secretion? 3. What is the role of HMGB1 in regulation of stretch of VSMCs? Reviewer #2: Dear Author, I have received manuscript entitled “PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cells; this is a good and significant article. In the present article author shown MS stressed VSMCs cells induced secreation hof HMGB1 via acetylation and phosphorylation of it. Also shown PDGFR-β pathways is critical for regulation of HMGB1 secretion during MS in VSMCs. Which further prove their hypothesis by knockdown and recombinant protein. This study is interesting, and manuscript can be considered for its publication. Nevertheless, I have few remarks concerning the manuscript. The manuscript can be considered for its publication after making the suggested changes. Comments. • NE-PERTM nuclear and cytoplasmic extraction, need to change with NE-PERTM • Isolation of Endothelial cells of the excised ---------. Need to more elaborate. Is any enzymatic method also used for isolation? • IP protocol need to elaborate. Also need to show pulldown with IgG as control. • HMGB1 ELISA kit (Elabscience, Houston, TX, USA) mentions cat no. also. • Statical analysis need to clearly mention about which group used ANOVA and in which group t test. • How about HMGB1 mRNA expression does it also change in MS . • Figure1 for mechanical stretch is not stimulation it should be cells stressed with Flexercell Tension Plus FX-4000T system. • Figure 2, Run cytoplasmic and nuclear fraction into same gel and show pattern of housekeeping genes lamin B and actin both fractions which confirm your any cross contamination of fractions. • Mention in each blot (Kda), size of proteins observed in immunoblot. • Figure 3.In the IP experiment of acetylated- and phosphorylated HMGB1- need to show IgG as a negative control also. • In the results section clearly mention after pulldown with HMGB1 immunoprobe with phospho or acetyl antibodies. • Figure 4 HMGB1 secretion via PDGFR signaling in MS-stimulated VSMC, Mention concentration used for all inhibitors either Method section or in results. • Figure 6, Knockdown efficiency does not clear, repeat the western and put clear significant change . Also add mRNA level of these genes in knockdown cells. Reviewer #3: Kim et al studied the mechanism of vascular smooth muscle cell (VSMC) dysfunction under mechanical stress. The authors isolated VSMCs from rat thoracic aorta and cultured. When the VSMCs were subjected to 3% mechanical stress, an increase in the time-dependent cellular release of high motility group box 1 (HMGB1), a major damage associated molecular pattern, was observed. The mechanical stress also augmented the cytosolic translocation of nuclear HMGB1. Mechanical stress induced releases of HMGB1 was mediated by platelet-derived growth factor receptor (PDGFR), especially by PDGFR-beta signaling. This study reported a novel mechanism of VMSC dysfunction. The manuscript is well written supporting a research experiment that was planned properly. The authors discussed that this finding will be helpful to manage conditions such as hypertension, but smaller resistance vessels more role in the development of hypertension. Is it possible to duplicate the finding using VSMCs from smaller resistance vessels? In the context of this experiment, interesting questions to be addressed by this group or any other group are: 1. whether AG1295, an inhibitor of PDGFR, can manage hypertension which is associated with mechanical stress of VSMCs? 2. What is the role of platelets under mechanical stress for the development of hypertension? ****** 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 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.0265191.r001
Revision 1
15 Dec 2021 Author Response Reviewer #1: In the present study, the authors studied signaling mediating release of HMGB1, a damage-associated molecular which mediates vascular complications in stress vasculature, in mechanically stressed vascular smooth muscle cells (VSMCs) isolated from mouse aorta SD rats. They found that HMGB1 released from stressed VSMCs was increased which accompanied with increased cytosolic translocation of nuclear HMGB1, and acetylated and phosphorylated form of HMGB1. Furthermore, they found that PDGFR inhibitor AG1295 inhibited HMGB1 release in stressed VSMCs. Moreover the increased HMGB1 release from stressed VSMCs was diminished when PDGFR-β was silenced by siRNA. PDGF-DD increased HMGB1 release. Finally the authors demonstrated that PDGFR-β-mediated secretion of HMGB1 in VSMCs might be a promising therapeutic strategy for hypertension-related vascular complications. The major concern of this manuscript is that the authors did not provide any evidence showing that the role of PDGFR-β/p-PDGFR-β-mediated HMGB1 in biology and function (i.e. contractile property) of VSMCs which makes the conclusion very weak. Other concerns including: � As we absolutely agree with your comments, the following contents were included in the revised manuscript on page 14: “HMGB1 is known as a multifunctional protein which induces vascular remodeling in hypertension via phenotypic transformation of VSMCs from contractile to synthetic type (REF #31, 32, 33).” 1. References should be added regarding the prerequisite of the cytosolic translation of nuclear HMGB1 for the secretion of HMGB1. � The translocation of HMGB1 during cell activation is mediated by acetylation and phosphorylation processes, and is prerequisite for the secretion of HMGB1 (REF #27). These contents were included in the result section of the revised manuscript on page 9-10. 2. How p-PDGFR-β regulates HMGB1 cytosolic translocation and secretion? � Interestingly, the MS-induced HMGB1 secretion was significantly attenuated in PDGFR-β-deficient cells, but not in cells transfected with siRNA for PDGFR-α. Likewise, PDGF-DD, but not PDGF-AA, directly increased HMGB1 secretion in VSMCs, indicating a pivotal role of PDGFR-β signaling on MS-induced HMGB1 secretion in VSMCs. Considering our results in which MS increased HMGB1 release via acetylation and phosphorylation of HMGB1, it is suggested that PDGF-β signal pathway plays a pivotal role in MS-induced HMGB1 acretylation and phosphorylation contributing VSMC release. These contents were included in the revised manuscript on page 15. 3. What is the role of HMGB1 in regulation of stretch of VSMCs? � HMGB1 is known as a phenotypic modulator of VSMCs from contractile to synthetic type, might attenuates contractile property of VSMCs (REF #31, 32, 33). These contents were included in the revised manuscript on page 14. Reviewer #2: Dear Author, I have received manuscript entitled “PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cells; this is a good and significant article. In the present article author shown MS stressed VSMCs cells induced secreation hof HMGB1 via acetylation and phosphorylation of it. Also shown PDGFR-β pathways is critical for regulation of HMGB1 secretion during MS in VSMCs. Which further prove their hypothesis by knockdown and recombinant protein. This study is interesting, and manuscript can be considered for its publication. Nevertheless, I have few remarks concerning the manuscript. The manuscript can be considered for its publication after making the suggested changes. Comments. • NE-PERTM nuclear and cytoplasmic extraction, need to change with NE-PERTM � The revised manuscript has been revised as your indications on Page 5. • Isolation of Endothelial cells of the excised ---------. Need to more elaborate. Is any enzymatic method also used for isolation? � In our primary culture process for VSMCs, endothelial cells were not isolated but just physically peeled off with stainless steel scraper, and any enzymatic method was not used. These contents were included in the revised manuscript on Page 6. • IP protocol need to elaborate. Also need to show pulldown with IgG as control. � To verify whether the antibody only affects target protein, HMGB1, we evaluated positive control (IP input) instead of negative control (IgG), as shown in other previous report by Xu et al. (Front Physiol. 2019, 5;10:854. doi: 10.3389/fphys.2019.00854). The procedures for IP were thoroughly revised in the revised manuscript on Page 7. • HMGB1 ELISA kit (Elabscience, Houston, TX, USA) mentions cat no. also. � Manuscript has been thoroughly revised as your indications. • Statical analysis need to clearly mention about which group used ANOVA and in which group t test. � Manuscript has been thoroughly revised as your indications. • How about HMGB1 mRNA expression does it also change in MS . � In our study, we investigated the regulatory signals involved HMGB1 secretion in MS-stimulated VSMCs, but not transcriptional regulation of HMGB1 synthesis as shown in other paper (Volchuk A et al. Nat Commun. 2020 Sep 11;11(1):4561. doi: 10.1038/s41467-020-18443-3). Thus, we focussed on the translocation and secretion of HMGB1 in VSMCs stimulated by MS. • Figure1 for mechanical stretch is not stimulation it should be cells stressed with Flexercell Tension Plus FX-4000T system. � Manuscript has been changed as your indications (Page 27). • Figure 2, Run cytoplasmic and nuclear fraction into same gel and show pattern of housekeeping genes lamin B and actin both fractions which confirm your any cross contamination of fractions. � Unfortunately, we followed the previous method showing sphingosine kinase 1-regulated translocation of HMGB1 by Tian T et al. (Cell Death Dis. 2020;11(12):1037. doi: 10.1038/s41419-020-03255-6u I). In the next study, we sincerely hope to run cytoplasmic and nuclear fraction into same gel. • Mention in each blot (Kda), size of proteins observed in immunoblot. � All of the figures have been revised as your indications. • Figure 3.In the IP experiment of acetylated- and phosphorylated HMGB1- need to show IgG as a negative control also. � To verify whether the antibody only affects target protein, HMGB1, we evaluated positive control (IP input) instead of negative control (IgG), as shown in other previous report by Xu et al. (Front Physiol. 2019, 5;10:854. doi: 10.3389/fphys.2019.00854). • In the results section clearly mention after pulldown with HMGB1 immunoprobe with phospho or acetyl antibodies. � Manuscript has been revised as your indications on Page 10. • Figure 4 HMGB1 secretion via PDGFR signaling in MS-stimulated VSMC, Mention concentration used for all inhibitors either Method section or in results. � The concentrations used for all inhibitors for Figure 4 were included in the figure legends in the revised manuscript on Page 28. • Figure 6, Knockdown efficiency does not clear, repeat the western and put clear significant change . Also add mRNA level of these genes in knockdown cells. � As we absolutely agree with your indications, Western blot data showing clear changes (Figure 6 in the revised manuscript) were included in the revised manuscript. Reviewer #3: Kim et al studied the mechanism of vascular smooth muscle cell (VSMC) dysfunction under mechanical stress. The authors isolated VSMCs from rat thoracic aorta and cultured. When the VSMCs were subjected to 3% mechanical stress, an increase in the time-dependent cellular release of high motility group box 1 (HMGB1), a major damage associated molecular pattern, was observed. The mechanical stress also augmented the cytosolic translocation of nuclear HMGB1. Mechanical stress induced releases of HMGB1 was mediated by platelet-derived growth factor receptor (PDGFR), especially by PDGFR-beta signaling. This study reported a novel mechanism of VMSC dysfunction. The manuscript is well written supporting a research experiment that was planned properly. The authors discussed that this finding will be helpful to manage conditions such as hypertension, but smaller resistance vessels more role in the development of hypertension. Is it possible to duplicate the finding using VSMCs from smaller resistance vessels? � Although VSMCs in smaller resistance vessels play important role in blood pressure regulation, we can hardly culture VSMCs from smaller resistance vessels. Thus, we could not perform an experiment using VSMCs in smaller resistance vessels. However, we think that there is high probability of the same result in a smaller resistance vessel. In the context of this experiment, interesting questions to be addressed by this group or any other group are: 1. whether AG1295, an inhibitor of PDGFR, can manage hypertension which is associated with mechanical stress of VSMCs? � In our present study, we investigated HMGB1 release in mechanically stressed VSMCs, but not contractile regulation of VSMCs. Considering the previous reports describing the role of HMGB1 on vascular complications, it is suggested that PDGFR inhibitor might be useful to prevent vascular complication caused by hypertension. 2. What is the role of platelets under mechanical stress for the development of hypertension? � Considering the previous reports describing the role of platelet activation in hypertension (Griffin G, Am Fam Physician, 2005;71(5):897-9; Gkaliagkousi E et al, Am J Hypertens, 2010;23(3):229-36; El Haouari M et al, Blood Cells Mol Dis, 2009;42(1):38-43), it is suggested that platelet might play an important role under mechanical stress for the development of hypertension. Considering the facts that PDGF was released from platelets (Linder BL et al, Proc Natl Acad Sci U S A, 1979;76(8):4107-11), future experiments are needed to determine the active role of PDGF under mechanical stress for the development of hypertension. Attachments Attachment Submitted filename: Response to reviewer.docx https://doi.org/10.1371/journal.pone.0265191.r002
28 Feb 2022 Decision Letter - Suresh Kumar Verma, Editor PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cells PONE-D-21-21832R1 Dear Dr. Kim, 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, Suresh Kumar Verma, PhD 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 #2: All comments have been addressed Reviewer #4: 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 #2: Partly Reviewer #4: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: I Don't Know Reviewer #4: 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 #2: Yes Reviewer #4: 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 #2: (No Response) Reviewer #4: 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 #2: Title PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cells, Baek etal addressed my comments. Don't have any further Reviewer #4: In the revised manuscript, the authors have adequately addressed the reviewer's comments. It can be published from a scientific point of view. ****** 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 #2: No Reviewer #4: Yes:** Prabhat Ranjan https://doi.org/10.1371/journal.pone.0265191.r003
Formally Accepted
8 Mar 2022 Acceptance Letter - Suresh Kumar Verma, Editor PONE-D-21-21832R1 PDGFR-β signaling mediates HMGB1 release in mechanically stressed vascular smooth muscle cells Dear Dr. Kim: 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. Suresh Kumar Verma Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0265191.r004

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .