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Fig. 3 | BMC Cancer

Fig. 3

From: Berberine, a natural compound, suppresses Hedgehog signaling pathway activity and cancer growth

Fig. 3

BBR inhibits the Hh signaling pathway activity by potentially targeting Smo. a BBR had no effect on the Gli luciferase activity stimulated by ectopic expression of Gli2N. NIH-3 T3 cells transfected with Gli2N, Gli luciferase reporter and the Renilla-TK constructs were treated with GANT61 and various concentrations of BBR for 36 h. GFP was used as a control for Gli2N. The results are expressed as mean ± s.d. from three independent experiments (n = 3). b BBR exhibited no effect on the Gli luciferase activity provoked by knockdown of Sufu. NIH-3 T3 cells were infected with lenti-virus harboring Sufu shRNA or shRNA control (the inserts shows the efficacy of knockdown by western blot analysis). The cells were further transfected with Gli luciferase reporter and the Renilla-TK constructs, and exposed to GANT61 or BBR for 36 h. Data are expressed as means ± s.d. (n = 3). (c-d) BBR significantly inhibited the Gli luciferase activity provoked by ectopic expression of Smo (c), while exhibited no effect on that provoked by ectopic expression of SmoM2 (d). NIH-3 T3 cells were transfected with Smo or SmoM2, Gli luciferase reporter and the Renilla-TK constructs were treated with various concentrations of BBR. GFP were used as a control for Smo or SmoM2. The results are expressed as mean ± s.d. from three independent experiments (n = 3). e BODIPY-cyclopamine competition analysis. Photographs are representatives from three distinct experiments. 293 T cells transfected with hSMO expression construct were exposed to 1 μM BODIPY-cyclopamine supplemented with or without cyclopamine, itraconazole or BBR for 10 h. The cells were then mounted with DAPI and photographed. Photographs are representatives from three distinct experiments (f). BODIPY-cyclopamine bound to Smo in 293 T cells as monitored by FACS analysis. #p > 0.05

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