Fig. 2: Shh signaling promotes stemness and imparts Imatinib resistance.
A Cartoon representing cell line model used, K562 (in black) and Shh-K562 (in green). B Shh and Gli-1-mRNA expression in Shh-K562 determined using RT-PCR, normalized to K562. Bar represents mean ± SD from three biological repeats. Western blot demonstrating expression of Gli-1 and Shh proteins in K562 and Shh-K562. Actin is used as loading control. C Levels of stemness markers, CD73, CD34, CD117, and CD90, and myeloid differentiation marker (CD38) in the Shh-K562 cell line determined using RT-PCR, normalized with K562. Bar represents mean ± SD from two biological repeats. D Representative images of FACS-based analysis of CD90, CD34, and CD73 expression in K562 and Shh-K562. Experiment was done with two biological and technical repeats with similar results. E Concentration curve for Imatinib to identify IC50 at 72 h, Shh-K562 (green line) compared to K562 (control; black line graph). Line graph represents mean ± SD from three biological repeats. F Images demonstrating colony forming units (CFU) using K562, Imatinib-treated K562 (TK562), Shh-K562, and Imatinib-treated Shh-K562 (TShh-K562). Graph represents average area of the colonies (30–100 colonies per condition) from duplicate experiments, in Log10 scale. For all images, P values, *<0.05, **<0.01, and ***<0.001. Statistical analysis done using t-test.
