Hh inhibition by compounds 21 and 22. The dose-response curve in SAG-treated NIH3T3 Shh-Light II cells (A,B) or mouse embryonic fibroblasts (MEFs) transfected with 12XGliBS-Luc and pRL-TK Renilla (normalization control) plus control (empty) or Gli1 vector (C,D). Cells were treated with increasing concentrations of compounds 21 (A,C) and 22 (B,D). Treatment time was 48 h and 24 h for NIH3T3 Shh-Light II cells and transfected MEFs, respectively. Data were normalized against Renilla luciferase. Data show the mean ± SD of three independent experiments. (*) p < 0.05 vs. SAG or Dimethyl sulfoxide (DMSO); (**) p < 0.01 vs. SAG or DMSO.