Review Reports

Round 1

Reviewer 1 Report

The authors investigated the antitumor effect of the deubiquitylating enzyme in-30 inhibitor PR-619 in cisplatin-resistant bladder UC. The study can be improved by correcting figure legends and providing details in materials and methods section.

1. Figure 1B. Please list number of mice used in each group in the experiment
2. X axis description in Figure 1 C, D are not clear, missing saline group and comparison groups.
3. What is the p value for Figure 1D experiment?
4. Please list antibodies catalog numbers and concentrations used for both western blot and IHC in the material and methods.
5. List correct dose for Figure 3b. Figure legend - PR-619 (15 μM), results text - PR-619 (20 μM)
6. Figure 4 is missing a C panel; C panel is listed in figure legend.
7. Figure 4 is missing D panel; D panel is listed in results section
8. Please provide a better description on how IHC score were calculated; what was the intensity scale used
9. For western blots with multiple bands put an arrow pointing to the correct band location (phosphor-JNK, caspase-4)

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

In the manuscript ijms-1413570, Huang et al. investigated the antitumor effect of DUB inhibitor (PR-619) in cisplatin-resistant bladder urothelial carcinoma. The authors showed that PR-619 improved both apoptotic and cytotoxic effects of cisplatin in cisplatin-resistant T24/R cells, which was associated with concurrent suppression of c-Myc expression. In a xenograft nude mouse model, PR-619 significantly improved the antitumor effects of cisplatin. These results highlight the therapeutic target of DUB for efficient treatment of cisplatin-resistant urothelial carcinoma. Overall, this study is well-designed and performed. The methods and results are adequately described. Accordingly, I would recommend the publication of this study after addressing the following minor concerns;

• the animal experiment is not well detailed. What the no of mice? control? grouping?
• Why the authors decided to in vitro test PR-619 at 20uM? please include it in the main MS (better in line 144).
• I suggest that the authors extend their discussion part to cover the most recent studies about Cisplatin and connect these to their findings.
• The authors should discuss, what would be the effect of PR-619 on specific DBU? also what the author think about the effect of specific DBU inhibition on both Cisplatin effect and treatment of bladder cancer? 
• Please add a new section for conclusion and outlook. Lines 248-252 are not sufficient.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf