Fig. 6

MEKK2/3 inhibit Hh pathway-dependent medulloblastoma tumor cell proliferation. a qRT-PCR analysis of Daoy cells treated with Shh CM for indicated times. b Daoy cells were treated with cyclopamine (5 μM) for 12 h and subjected for qRT-PCR analysis. c Daoy cells stably expressing GLI1-WT, GLI1-6A, and GLI1-6D were subjected to cell proliferation assay using CCK8. d Cell lysates from GLI1-WT, GLI1-6A, and GLI1-6D Daoy cells were analyzed by western blot with indicated antibodies. e Daoy cells stably expressing GLI1-WT, GLI1-6A, and GLI1-6D were subcutaneously injected into the nude mice (five mice for each group), and tumor weights were measured 20 days later. f WT and DKO Daoy cells (5 × 10⁶) were subcutaneously injected into nude mice (six mice for each group), and tumor volume was measured and calculated. g Hela cells were transduced with MEKK3 lentivirus and subjected to cell proliferation assay using CCK8. h Cell lysates from g were analyzed by western blot. i Heat map analysis reveals that the expression of MEKK2/3 is negatively correlated with Hh pathway activity in GSE37418 dataset. j Daoy cells were transduced with indicated lentivirus and subjected to colony formation assay. k The diagram shows that activated MEKK2/3 by FGF2 directly phosphorylate GLI1. Phosphorylation results in reduced GLI1 protein stability, transcriptional activity, and tumorigenesis potency. *P < 0.05, **P < 0.01, and ***P < 0.001 (two-tailed Student’s t-test). Quantitative data were presented as mean ± s.d. from a representative of at least three independent experiments