Fig. 1

MINDY1 depletion decreases Hippo signaling activity in bladder cancer cells. A The siRNAs specific to each deubiquitinating enzyme (MINDY1-4) were transfected into T24 cells. After 48 h, cells were lysed and the YAP protein level was analyzed by Western blot. Relative YAP protein level was normalized to GAPDH. B MINDY1 depletion decreased YAP protein level. C MINDY1 depletion did not affect YAP mRNA level. D Genetic alternations of MINDYs in bladder cancer. E, F MINDY1 depletion decreased YAP target genes. Bladder cancer cells were transfected with si MINDY1 or siControl. Total RNA was prepared and the expression of the endogenous YAP target genes, CTGF, CYR61 and ANKRD1 were determined by qRT-PCR. G, H MINDY1 depletion affected YAP/TEAD-luciferase activity. Bladder cancer cells were transfected with siMINDY1 or siControl together with YAP/TEAD luciferase reporter plasmid. Luciferase activity was measured 48 h after transfection. *, P value < 0.05; **, P value < 0.01; ***, P value < 0.001