Figure 3

Pharmacologic and lentiviral SMO inhibition inhibits downstream Hh signaling in CP-CML cells.

(A) Expression of Gli1 in total BM cells, BM MPP and LTHSC subpopulations and in BM stromal cells from Bcr-Abl-expressing mice (n = 8 per arm) treated with LDE225, nilotinib, LDE225 + nilotinib or vehicle alone for 5 days. Results represent the mean ± SEM for multiple samples. (B) Expression of GLI1 following 72h exposure to LDE225 in CD34⁺ CP-CML cells (n = 7). Results represent the mean ± SEM for multiple samples. (C,D) Expression of targets and mediators of Hh signaling in CD34⁺ CP-CML cells following shRNA mediated SMO KD; (C) Representative flow cytometry histogram demonstrating comparative transfection efficiency of lentiviral vector containing GFP-SMO KD shRNA or GFP-Scram shRNA compared to untransfected cells. (D) Expression of SMO (Di), GLI1 (Dii) and the Hh pathway inhibitor GLI3 (Diii) in CD34⁺ CP-CML cells following shRNA mediated SMO KD normalised to GAPDH and compared to the scrambled control. Results shown represent the mean ± SEM for 3 independent transfections of different CD34⁺ CP-CML samples. Significance values; *p < 0.05; **p < 0.01; ***p < 0.005.