Peer-reviewed by 2 reviewers with median rating of 15/20. Review process was triple-blinded.
Round 1 (14/20)
Round 2 (15/20)
Conceptual advance and Impact7
The findings are novel and somewhat unexpected, and the study adds to the body of knowledge in the regulation of viral-mediated tumorigenesis.
In the final sentence, the importance of the results may be overstated because interactions with full length LT were already characterized. If the authors agree, it could be possible to change part of the final sentence to say 'with the Brd4 protein add to existing knowledge about interactions with LT and its role in tumorigenesis, and assist in efforts to more precisely define new therapy targets for this disease''
Suggested spelling/grammatical corrections:
'to transcriptionally regulate the'
'of virus-associated cancers'
'showed enhanced transcriptional activity, whereas'
Results & Discussion
Good results section with suggested major and minor corrections or clarifications:
Explain or speculate why differences in binding by the three different Abs occurs in Figure A in the MKL1 cells compared to the MS1 cells, and also why the Abs appear to be binding a different MW target between the two cell lines.
Explain the differences in the rows of images of Fig B. Are they replicates? They appear similar.
Minor requests or suggested changes:
'by gentle pipetting to obtain a single cell suspension that adhered to....'
Specify whether the 'range 4.4 to 11.9' relates to the percentage or the correlation coefficient.
Specify what 7.8% is referring to.
Add a space in between sT and alone (currently reads sTalone).
To assist others in replicating the immunofluorescence assay, can you please indicate if (and how) you washed the EDTA-treated cells before incubating on lysine coverslips?
Please specify that you used truncated LT in the title of the study
Specify what the different rows signify in Fig B.
Indicate the number of experimental repeats yielding similar results for each result.
Minor suggested requests:
Specify the technique used to isolate nuclear proteins in the first sentence of the legend.
Specify whether the error bars are SD or SEM.
Use an acronym or the word empty instead of '-' to designate the empty lanes Figure A and the legend.
** usually signifies p<0.01, so please verify the value stated in the figure legend for figure C and in the results section is correct, and adopt accepted standards of asterisk notation of statistical significance.
Unclear what LT168 is and why it is included...the figure legend alone should allow a reader to interpret the results (with this in mind it would be helpful to add a few extra labels to figures A, B, and D where appropriate).
The results presented in the study do not specifically demonstrate whether there is viral replication or not, so the statement 'in the absence of virus replication', even though likely to be correct based on prior knowledge, should probably be omitted from the conclusion.
Please leave a space in between the words LT and antigen (currently reads LTantigen)
unusual sentence, potentially missing a word:
'with the same ?antibodies? was challenging'
For completeness, and if you believe it is the case, can you please include a statement somewhere in the manuscript to confirm that the overall findings are not due to different binding abilities of the 3 Abs in the different assays?
Conceptual advance and Impact5
Interesting observations that can lead to a better understanding of the function of truncated LT in MCC development.
"transcriptionally regulates" should be "transcriptionally regulate".
LT is suddenly an abbreviation for Large T but this abbreviation is never introduced. perhaps it also should be added MCV LT not just any LT.
BRD4 not Brd4.
"was to validate this interaction"? Which interaction? I guess BRD4-LT?
Results & Discussion
"did co-localize with BRD4 protein in nucleus, although weakly at only at 7.8 % with a correlation coefficient of 0.36 (range 4.4 to 11.9)": This shoudl be better explained, perhaps supported by arrows in the figure where BRD4 and LT4 supposedly are co-localized. What does 7.4% mean? 7.8% of all LT or of all BRD4 interacts with the other protein? How does this relate to the Western blots in (A)?
the term "microbe" is used in an unusual way. I would not have used it in this context.
BRD4 and Brd4 are used without logic. I guess when talking about human BRD4 protein it should be all capitals.
"drive tumorigenesis in Merkel cells": not sure this is correct. since Merkel cells seem highly differentiated, the target may be certain keratinocytes from which Merkel cells are derived from?
McKinney not Mckinney