Fig. 1

BBR inhibits Hh pathway activity in vitro. a Chemical structure of BBR. b BBR inhibited the Gli luciferase activity provoked by ShhN CM and SAG. NIH-3 T3 cells transfected with luciferase reporter plasmids containing 8 × Gli binding sites and Renilla-TK plasmids were exposed to ShhN CM or SAG supplemented with various concentrations of BBR for 36 h. The cells were lysed for the dual luciferase reporter activity analysis. The firefly luciferase activity was normalized by the Renilla luciferase activity. The results are expressed as means ± s.d. from three independent experiments (n = 3). (c-d) BBR inhibited the mRNA expressions of Gli1 provoked by ShhN CM (c) and SAG (d). NIH-3 T3 cells were treated with ShhN CM or SAG with or without various concentrations of BBR for 24 h and harvested for RT-qPCR analysis. Data are expressed as mean ± s.d. (n = 3). Experiments were repeated for three times. e BBR inhibited alkaline phosphatase activity in C3H10T1/2 cells. C3H10T1/2 cells were treated with or without ShhN CM supplemented with various concentrations of BBR for 72 h. The results are expressed as mean ± s.d. from three independent experiments (n = 3)