All reviews of published articles are made public. This includes manuscript files, peer review comments, author rebuttals and revised materials. Note: This was optional for articles submitted before 13 February 2023.
Peer reviewers are encouraged (but not required) to provide their names to the authors when submitting their peer review. If they agree to provide their name, then their personal profile page will reflect a public acknowledgment that they performed a review (even if the article is rejected). If the article is accepted, then reviewers who provided their name will be associated with the article itself.
All remaining concerns of the reviewer were adequately addressed, and revised manuscript is acceptable now.
[# PeerJ Staff Note - this decision was reviewed and approved by Gwyn Gould, a PeerJ Section Editor covering this Section #]
My concerns have been adequately addressed
My concerns have been adequately addressed
My concerns have been adequately addressed
Please address the remaining concerns of reviewer #1 and amend the manuscript in line with the proposed changes.
Figures 1 and 2 are a bit better, but still difficult to read. Font size of all writing (legends, axes, labels, etc.) should be increased so that it is legible when printed out.
Methods detail is significantly improved. However, the authors do not address the issue that sepsis drives cell death and therefore protein normalization may not be sufficient in lieu of a live cell count. An increase in dead cells, which still contain protein, may artificially diminish respiration measurements. Authors should normalize to CCK-8, crystal violet, or trypan blue exclusion counts, or at the very least note this as a weakness in the interpretation of the data. Also, please define the seahorse data calculations within the body of the methods (e.g. state directly that ATP production was determined as the difference in O2 consumption before and after oligomycin, etc.)
Given that the authors state in their response that they do not observe a significant difference in ATP, lactate,or any OCR values between control and nucleolin-silenced cells in the presence of LPS and TNFa i, the statements on lines 270-273 “and also aggravated the decrease in basal respiratory level, reserve respiratory capacity, maximal respiratory capacity, and ATP production capacity after stimulation(Figure 4F-I), suggesting that down-regulated nucleolin aggravated the decrease in the overall oxidative phosphorylation level of the cells after TNF-α combined with LPS stimulation” is not supported by the data at this time. The fact that no significant difference between these groups must be clearly stated in the manuscript and discussed. Further, the section 3.4 heading (lines 256-257) is misleading, since metabolism is not significantly altered by nucleolin deficiency during combined stimulation, only in the non-stimulated control groups. Please correct these statements, or repeat the experiments with a sufficient sample size to support the supposition.
No comment
No comment
No comment
All my queries were answered satisfactorily. I would recommend the manuscript for acceptance.
Please address the concerns of both reviewers and amend the manuscript accordingly.
**PeerJ Staff Note:** Please ensure that all review and editorial comments are addressed in a response letter and that any edits or clarifications mentioned in the letter are also inserted into the revised manuscript where appropriate.
**Language Note:** The review process has identified that the English language must be improved. PeerJ can provide language editing services - please contact us at [email protected] for pricing (be sure to provide your manuscript number and title). Alternatively, you should make your own arrangements to improve the language quality and provide details in your response letter. – PeerJ Staff
Although the authors have clearly submitted a large number of findings in a very interesting mouse model in figures 1 and 2, the fonts used are very difficult to read and therefore these figures are hard to assess. In addition, the English should be reviewed for grammar and syntax errors.
How were ATP and lactate measurements normalized? Given that sepsis results in cell death, results should be normalized to living cell counts. In addition, please provide more data for the seahorse methods. What reagents and concentrations are used for each stage, how are results normalized, and how are basal, spare capacity and atp production determined? The O2 consumption methods description appears to describe the use of animal subjects, but only data from H9c2 cells is shown.
In figure 4, it is unclear whether the two treatment groups (LPS and TNFa) produce a different effect in the overexpressor or silenced cell groups compared to control. Since the authors’ thesis is that nucleolin impacts the effect of this treatment on ATP production, lactate levels and respiration, this comparison is key. For the seahorse data, by eye it appears that this is the case, although it it not indicated by statistical notation. Please complete these comparisons and add them to the figures. In addition, it is difficult to assess the findings of figures 1 and 2 due to legibility.
The use of the English language in this manuscript is professional and clear. Provided literature references are generally sufficient, and the figures are well-labeled and described. However, I’d recommend improving the quality and resolution of some figures to enhance readability.
The materials and methods section is adequately described.
The manuscript demonstrates originality, accompanied by a well-defined problem statement and a clear focus on addressing a gap within the field of study. Although the presented data is sufficiently robust, minor edits in several places are advisable for further enhancement.
I provided some corrections and a few queries to enhance the quality of the manuscript.
All text and materials provided via this peer-review history page are made available under a Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.