Fig. 7.

Melatonin inhibits USP10-mediated HDAC7 deubiquitination thus inducing HDAC7 proteolysis. a Representative IHC images for USP10 expression in ESCC tissues. Scale bar, 200 μm and 50 μm (inset), respectively. b Statistical analysis of USP10 expression in 148 ESCC patients through IHC staining. c Kaplan–Meier survival analysis by high/low USP10 levels of the 148 ESCC patients based on the microarray tissue IHC results. d The spearman correlation analyses of USP10, HDAC7 and c-Myc expression in the ESCC tissues. e Kaplan–Meier analysis of the association between overall survival and the expression of USP10, HDAC7 and c-Myc in 148 ESCC patients. f Photographs and results showing xenograft tumor morphologies and weights in each group after subcutaneously injection of shCon or shUSP10 EC109 cells, respectively. g Representative images and results of colony formation and EdU incorporation assays in the EC109 cells with melatonin and USP10 knockdown treatment. h Western blotting results showed melatonin treatment for 48 h decreased USP10 levels in ESCC cells. i Immunoblot analyses of input and HDAC7 immunoprecipitated from EC109 cells with melatonin exposure and treated with 10 μmol/L MG132 overnight before harvesting. j Representative western blotting results of USP10 and HDAC7 in the EC109 cells with melatonin and USP10 knockdown treatment. ^*P < 0.05 vs. the shCon group, ^#P < 0.05 vs. the shUSP10 #1 group, ^&P < 0.05 vs. the USP10 #2 group, ^$P < 0.05 vs. the MEL 4 mmol/L group. USP10 ubiquitin-specific protease 10, HDAC7 histone deacetylase 7, ESCC esophageal squamous cell carcinoma, ub ubiquitin, MEL melatonin, LV lentivirus, IHC immunohistochemistry, USP10 ubiquitin-specific peptidase 10