Fig. 4: USP12 promotes Hippo/YAP signaling in gastric cancer.

A Western blotting analysis showing USP12 depletion decreases YAP protein stability. AGS and MKN74 cells transfected with 50 nM siControl or two independent siUSP12. Cell lysates were immunoblotted with the indicated antibodies. β-Actin was used as internal control. B, C qRT-PCR analysis of YAP target genes (CTGF, CYR61) expression. AGS and MKN74 cells transfected with 50 nM siControl or two independent siUSP12 for 48 h. Total RNA was extracted for gene expression analysis. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for target gene expression comparison. Luciferase assays showing USP12 depletion affects TEAD-luciferase activity in AGS (D) and MKN74 (E) cells. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for luciferase activity comparison. F Western blotting analysis showing USP12^WT increases YAP protein stability but not USP12^C48S. AGS cells were transfected with 1 μg Flag or Flag-USP12^WT/Flag-USP12^C48S. Cell lysates were immunoblotted with the indicated antibodies. β-Actin was used as internal control. G qRT-PCR analysis of YAP target genes (CTGF, CYR61) expression. AGS cells were transfected with 1 μg Flag or Flag-USP12^WT/Flag-USP12^C48S for 48 h. Total RNA was extracted for gene expression analysis. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for target gene expression comparison. H Luciferase assays showing USP12^WT increases TEAD-luciferase activity but not USP12^C48S in AGS cells. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for luciferase activity comparison.