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Fig. 6 | Military Medical Research

Fig. 6

From: Melatonin inhibits ESCC tumor growth by mitigating the HDAC7/β-catenin/c-Myc positive feedback loop and suppressing the USP10-maintained HDAC7 protein stability

Fig. 6

USP10 deubiquitinates and stabilizes HDAC7 protein in ESCC cells. a Indicated Flag-tagged USPs and HA-HDAC7 were transfected into HEK-293T cells, and Flag-tagged HDAC7 was respectively transfected into HEK-293T and EC109 cells, then co-IP analysis was performed to determine the interaction between HDAC7 and USP10 in vitro by Western blotting analyses. b Immunoblot analyses of whole cell lyses derived from HEK-293T cells transfected with indicated Flag-USP10, HA-HDAC7 and GFP plasmids. c Immunoblot analyses of input and HDAC7 immunoprecipitated from EC109 cells with USP10 knocking down, and treated with 10 μmol/L MG132 overnight before harvesting. d Result of HDAC7 mRNA levels in the EC109 cells with USP10 knocking down. e HEK-293T cells were transfected with indicated Flag-USP10 and HA-HDAC7 plasmids, then cells were treated with CHX at indicated time points. The HDAC7 protein level was quantified by the ImageJ software and plotted. f Overview of USP10 protein structures. g HEK-293T cells transfected with the indicated constructs were subjected to pull down with anti-HA or anti-GST. h HDAC7 ubiquitylation was analyzed in HEK-293T cells transfected with USP10/WT or USP10/C424A. USPs ubiquitin-specific proteases, HDAC7 histone deacetylase 7, ub ubiquitin, EV empty vector, WT wild type, CHX cycloheximide, USP10 ubiquitin-specific peptidase 10

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