PONE-D-21-32691Deletion of the scavenger receptor Scarb1 in osteoblast progenitors does not affect bone massPLOS ONE

Dear Dr. Ambrogini,

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PLOS ONE

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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

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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

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Reviewer #1: Yes

Reviewer #2: No

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Reviewer #1: Yes

Reviewer #2: Yes

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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 manuscript describes studies of the phenotype about Scarb1 conditional osteoblast-specific knockout mice using Osx1-Cre mice. The authors do not explain the phenomena that this conditional KO mice exhibited normal bone mass and analyze the reason of the dissociation between in vitro and in vivo experimentally. It is little of rationale. Unfortunately, I cannot recommend this paper to publish in PLoS One.

Reviewer #2: In this study, the authors investigated specific deletion of Scarb1 in bone marrow osteoblast lineage on bone mass of male and female mice.

Based on the findings that no differences in trabecular or cortical bone mass between the controls and Scarb1(fl/fl) in femur or spine, authors concluded that Scarb1 in osteoblast lineage does not play an important role in bone homeostasis and is not essential for the effects of PC-OxPL on osteoblasts (lines 39-40), or Scarb1 is not needed for osteoblast differentiation and function (lines 85-86).

As we know, bone mass reflects the balance of formation and resorption or the number and activity of osteoblasts and osteoclasts. Neither osteoblast nor osteoclast function was evaluated in the study.

Calvaria- or bone marrow-derived osteoblasts from these mice could be cultured to evaluate osteoblast function. Alternatively, osteoblast and osteoclast in bone tissue from these mice can be examined.

Line 220, the method is lacking on how vehicle or OxLDL was given

Figure 1, why only measure mRNA of osteocalcin and ALP? Were osteoblast colony, bone nodule, ALP staining performed on calvaria- or bone marrow osteoblast cultures?

Figure 1, data presented in the figure are different with the cited reference (lines 175-176) describing relative gene expression using 2(-Delta Delta C(T)) Method. Please provide raw CT data of either osteocalcin or ALP including the calculation of relative mRNA.

Lines 115-121 and Figure 2, instead of showing gDNA, can you show genotypes in gel as shown in page 4 of this paper to confirm deletion of Scarb1? Genomic DNA from the surfaces of bone doesn’t mean from osteoblasts only.

(Akiyama TE, Sakai S, Lambert G, Nicol CJ, Matsusue K, Pimprale S, Lee YH, Ricote M, Glass CK, Brewer HB, Jr., et al. Conditional disruption of the peroxisome proliferator-activated receptor gamma gene in mice results in lowered expression of ABCA1, ABCG1, and apoE in macrophages and reduced cholesterol efflux. Mol Cell Biol 2002;22(8):2607-19. doi: 10.1128/mcb.22.8.2607-2619.2002.)

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Reviewer #1: No

Reviewer #2: Yes: Jay Cao

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Deletion of the scavenger receptor Scarb1 in osteoblast progenitors does not affect bone mass

PONE-D-21-32691R1

Dear Dr. Ambrogini,

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,

Xing-Ming Shi, Ph.D

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

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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: (No Response)

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

Reviewer #2: (No Response)

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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: (No Response)

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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)

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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: (No Response)

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Reviewer #2: No