Fig. 4

Relationship between LGR5 and EMT and the Wnt/β-catenin pathway, and Wnt-C59 treatment in GSCs. a Images and statistical results in invasion assays (P < 0.001, n = 3; P < 0.001, n = 3, Student t test for shLGR5 and Lenti-LGR5, respectively) and migration assays (P < 0.001, n = 3; P < 0.01, n = 3, Student t test for shLGR5 and Lenti-LGR5, respectively) for transfected U251 GSCs. b Images and statistical results in invasion assays (P < 0.001, n = 3; P < 0.01, n = 3, Student t test for shLGR5 and Lenti-LGR5, respectively) and migration assays (P < 0.01, n = 3; P < 0.01, n = 3, Student t test for shLGR5 and Lenti-LGR5, respectively) for transfected 8591 GSCs. c, d Western blot analyses of transfected U251 and 8591 GSCs for LGR5, EMT related genes and surrogate markers for the activated Wnt/β-catenin pathway. e Images and statistical results of invasion assays (P < 0.001, n = 3; P < 0.001, n = 3; P < 0.05, n = 3, one-way analysis of variance (ANOVA) for 0 μΜ and 5 μM, 0 μΜ and 20 μM and 5 μM and 20 μM, respectively) and migration assays (P < 0.001, n = 3; P < 0.001, n = 3; P < 0.01, n = 3, one-way analysis of variance (ANOVA) for 0 μΜ and 5 μM, 0 μΜ and 20 μM and 5 μM and 20 μM, respectively) at 24 h after treated with Wnt-C59 in U251-GSCs. f Western blot analyses of U251 GSCs for LGR5, EMT related genes and surrogate markers for the activated Wnt/β-catenin pathway at 48 h after treated with Wnt-C59. g Images and statistical results of invasion assays (P < 0.01, n = 3; P < 0.001, n = 3; P < 0.05, n = 3, one-way analysis of variance (ANOVA) for 0 μΜ and 5 μM, 0 μΜ and 20 μM and 5 μM and 20 μM, respectively) and migration assays (P < 0.01, n = 3; P < 0.001, n = 3; P < 0.01, n = 3, one-way analysis of variance (ANOVA) for 0 μΜ and 5 μM, 0 μΜ and 20 μM and 5 μM and 20 μM, respectively) at 24 h after treated with Wnt-C59 in 8591-GSCs. h Western blot analyses of 8591 GSCs for LGR5, EMT related genes and surrogate markers for the activated Wnt/β-catenin pathway at 48 h after treated with Wnt-C59. Scale bar = 100 μm. All bars are represented as mean ± SD from triplicate wells. ***, P < 0.001; **, P < 0.01; *, P < 0.05