SMO binding assay. HEK293T cells were transfected with a SMO-expressing plasmid. After 48 h, the cells were fixed and incubated with BODIPYcyclopamine (green, 5 nM) and treated with either DMSO, vismo or A23 for 4 h. The nuclei were visualized by staining the cells with 4,6-diamidino-2-phenylindole (DAPI) (blue). The images are representative of three biological replicates ( n = 3). Scale bar, 30 m. For c e , statistical analyses were performed relative to DMSO/purmorphamine by employing unpaired two-tailed t -tests with Welch’s correction (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; NS, not significant).
