Figure 3.

Effects of SMO and/or PI3K inhibition on HH target gene expression, proliferation and apoptosis in human ERMS cell lines. RD and RUCH-2 cells treated with 10 μM of SMO inhibitors and/or 10 μM of the PI3K inhibitor pictilisib. (A) GLI1 expression levels after treatment for 24 h. HHIP expression was investigated after treatment with vismodegib and/or pictilisib (B) BrdU incorporation after treatment for 24 h. BrdU incorporation of solvent treated cells was set to 100%. Bars represent the mean + s.e. of three independent experiments performed in triplicates. (C) Annexin V staining and subsequent FACS analysis of RD cells treated for 48 h with the drugs as indicated. Bars represent the mean number of Annexin V⁺ cells + s.e. of two independent experiments performed in duplicates.^*P < 0.05, ^**P < 0.01, ^***P < 0.01 compared to cells treated with solvent and analyzed by Tukey's test for multiple comparisons. ^#P < 0.05 compared to cells treated with either drug alone.