FIGURE 3.

HH/GLI1 activation and suppression of AMPK in Vismodegib‐resistant medulloblastoma cell lines. (A) To generate Vismodegib‐resistant MB cells, MB11 and DAOY cells were treated with Vismodegib daily with a gradual increase in concentration from 10 to 300 μM in 16 weeks. (B) From two pairs of MB parental and Vismodegib‐resistant cell lines (VisR), MB11 and DAOY, mRNA was collected, and the amount of HH/GLI1 downstream targets, GLI1, PTCH1, Cyclin D1, C‐MYC, and BCL‐2 mRNA was analyzed using qRT‐PCR with GAPDH mRNA as the internal control for normalization. The bars indicate mRNA level relative to that of parental cells. The experimental points were in triplicate and independently repeated three times (*p < 0.05, **p < 0.01, *** p < 0.001). (C) Lysates from MB11 and DAOY (P: parental; VisR: Vismodegib‐resistant) cells were analyzed by immunoblot with the indicated antibodies