Fig. 7.

The failure of Islr-deficient myoblast differentiation is rescued by canonical Wnt signaling activation. a Western blot analysis of Axin1 protein levels in shCtrl and shIslr C2C12 cells incubated with Wnt3a or saline for 24 h after 2 d in differentiation medium. b Western blot analysis of active β-catenin protein levels in nuclear lysates extracted from shCtrl and shIslr C2C12 cells incubated with Wnt3a or saline for 24 h after 2 d in differentiation medium. c Intramuscular injection of Wnt3a at 1.5 d postinjury and Western blot analysis of Axin1 protein levels in injured TA muscles of control and Islr cKO mice at 4 d postinjury. d Intramuscular injection of Wnt3a at 1.5 d postinjury and Western blot analysis of active β-catenin protein levels in injured TA muscles of control and Islr cKO mice at 4 d postinjury. e Western blot analysis of Axin1 and GSK3β protein levels in shIslr C2C12 cells incubated with 1-AKP or DMSO after 7 d in differentiation medium. f Immunofluorescence staining for MyHC in shIslr C2C12 cells treated with 1-AKP or DMSO after 7 d in differentiation medium. N = 3 cell cultures in each group. The percentages of MyHC⁺ cells are shown on the right. g Immunofluorescence staining for MyoG in shCtrl and shIslr C2C12 cells treated with 1-AKP or DMSO after 3 d in differentiation medium. N = 3 cell cultures in each group. The percentages of MyoG⁺ cells are shown on the right. h Immunofluorescence staining for MyoG in shCtrl and shIslr C2C12 cells treated with CHIR or DMSO after 3 d in differentiation medium. N = 3 cell cultures in each group. The percentages of MyoG⁺ cells are shown on the right. i Immunofluorescence staining for MyoG in primary myoblasts of control and Islr cKO mice treated with CHIR or DMSO after 1 d in differentiation medium. N = 3 cell cultures in each group. The percentages of MyoG⁺ cells are shown on the right. Scale bars are all 50 μm. Error bars represent the means ± s.d. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001; Student’s t test