Fig. 1

SNS-032 inhibits cellular growth through blocking YAP signaling in uveal melanoma (UM) cells. a The protein levels of CDK7/9 in UM cells and ARPE-19 cells were examined by Western blot analysis. b SNS-032 inhibited CTD phosphorylation of RNA Pol II in UM cells. Cells were treated with increasing concentration of SNS-032 for 48 h, Western blot analysis with the indicated antibodies was performed. c SNS-032 blocked YAP signaling in UM cells. Mel270 and Omm2.3 cells were treated with SNS-032 for 48 h, the protein levels of YAP, phospho-YAP (S127) and its downstream targets CTGF and CYR61 were examined by Western blot analysis. d SNS-032 inhibited gene transcription of YAP. The UM cells were treated with SNS-032 for 24 h, the mRNA levels of YAP were detected by qRT-PCR analysis. e SNS-032 suppressed YAP-mediated gene transcription. UM cells were co-transfected with Gal4BD-TEAD4, G5-luciferase reporter (contains five Gal4-binding sites), and internal control Renilla luciferase reporter constructs for 24 h, then treated with SNS-032 for another 24 h, followed by luciferase activity assay. f SNS-032 reduced mRNA levels of YAP target genes. The UM cells were treated with SNS-032 for 24 h, qRT-PCR analysis of YAP target genes CTGF and CYR61 were performed. g SNS-032 inhibited cell viability of UM cells but not ARPE-19 cells. The cells were incubated with increasing concentrations of SNS-032 for 72 h, cell viability was measured by MTS assay. Dose-response curves from three independent experiments are shown. Error bars represent standard deviation (SD). h Anchorage-independent colony growth of UM cells was dramatically inhibited by SNS-032. The UM cells pretreated with different concentrations of SNS-032 were cultured in soft agar, colony-formation ability was assessed. i and j Ectopic expression of YAP attenuated the growth inhibition effect of SNS-032. UM cells stably expressing of YAP were treated with SNS-032 (0.5 μM) for the indicated durations, expression of YAP in UM cells was examined by Western blot analysis (i). The cell number was counted by trypan blue exclusion assay (j). *, P < 0.05; **, P < 0.01; ***, P < 0.001, one-way ANOVA, post hoc intergroup comparisons, Tukey’s test