N-Myc and STAT Interactor is an Endometriosis Suppressor
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript by Park, Guan, and Han titled “N-Myc and STAT interactor is an endometriosis suppressor” is a well written and laid out paper examining the role of NMI in endometriosis. Both a potential mechanism for the downregulation of NMI via HDAC8 as well as the role of NMI in blocking IFNA-induced cell death are examined. The experiments are well laid out and described. The overall conclusion of NMI having a role in suppressing endometriosis is supported. There are a couple points that need clarity for publication.
How does the ER-beta binding to the NMI promoter shown in Figure 2C differ between normal and endometriosis tissue? Yes, NMI binds, but is there really a difference between the 2 biological states?
What are the levels of IFNA found in endometriosis patient samples and how do they compare to the levels used in Figure 3? Are the 500 and 1000 units/mL physiologically relevant to endometriosis or are much lower amounts more relevant? This has significant implication for the RNA-seq analysis in Figure 4 and beyond as the data shown that 500 units has little effect, and 1000 units is needed.
It is not clear what the source of IFNA is in endometriosis. What cell(s) are the source and what levels of IFNA are present in the luciferase mouse model shown in Figure8? Given the need to use NOD-SCID mice with the human cell line models, how does the lack of an immune response to the injected cells alter the interpretation of the data?
A simple student’s t-test is not adequate for analysis of data with more than 2 groups. An ANOVA analysis with a pot-hoc correction for multiple comparisons or for multiple comparisons to a single control is required.
Minor Points
Quantification of the decrease in NMI levels with myc-tagged ER-beta overexpression in Figure 2 b is needed as the decrease is not clear from the provided blots.
The labels in Figure 6 are too small to read and need to be enlarged – specifically, the fold changes under the western blots.
Line 561 references Figure 9 that is not in the manuscript. Is a model figure missing as this is what seems to be referenced? The addition of a model figure would be very helpful for the reader.
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors present a fascinating article in which the possible role of the interaction between N-MYC and STAT as a therapeutic target for the treatment of endometriosis is evidenced because it shows how these two proteins act as suppressors of the adhesion and proliferation of endometriotic cells. Below are my comments to improve the quality of the manuscript.
1. Lane 76: Replace “progression. [16, 23].” with “progression [16, 23].”
2. Lane 82: Replace “…endometriosis [24] [15].” with “…endometriosis [15].”. Track change is evident.
3. Lane 109: Replace “…normal endometrium [32] [14].” with ““…normal endometrium [14].” Track change.
4. Figure 1A: Check the numbers that denote each of the blot lanes.
5. Figure 1A-C: The names should be consistent because both Figure 1B and 1B are quantifying the results shown in Figure 1A. Given the above, they should name Endometriotic or endometriosis, but not both.
6. Lane 154: Replace “… (Figure 2A) [31] [15].” with “… (Figure 2A) [15].” Track change.
7. Figures: statistics, authors must be consistent when showing information regarding statistics. In some graphics, they use *, **, or ***. However, in other graphs, they use the P-value.
8. Material and methods, statistics: how many mice were used, what was the n, and how was it calculated?
9. Material and methods, statistics: what was the n for the cell cultures? How was it calculated? Were there replicas? Although this is mentioned in some figures, it is part of the methods and should be made explicit here.
10. Material and methods, what is the ethical approval number or code for the study?
11. Lane 760: “… 23.1.rc1(Community, 2022).” with “… 23.1.rc1 (Community, 2022).”
12. Material and methods, statistical analysis: were all the data normally distributed? If not, what statistical tests were used?
13. Material and methods, statistical analysis: how were the analyses shown in the heatmap (Figure 4C) performed?
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe authors have adequately responded to the raised critiques.
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors did an excellent job responding to my previous concerns.
