Figure 5
From: Canonical Wnt signaling induces BMP-4 to specify slow myofibrogenesis of fetal myoblasts
Wnt-3a induces slow myosin heavy chain expression in fetal myoblasts via BMP-4. Fetal myoblasts were isolated by fluorescence-activated cell sorting (FACS) from E14.5-15.5 embryos of Myf5-Cre/Rosa26-YFP mice. FACS isolated fetal myoblasts were cultured and induced to differentiate for three days. (A-D) Immunofluorescence showing slow myosin heavy chain (green) expression in myotubes treated with vehicle control medium or Wnt-3a recombinant protein (50 ng/ml). (E) Relative expression of myosin heavy chain and BMP-4 genes based on qPCR analysis. (F) Western blotting showing slow myosin heavy chain protein expression. (G-J) Immunofluorescence showing slow- (green) and pan- (red) myosin heavy chain expression in myotubes treated with vehicle control medium, Wnt-3a, and BMP-4 recombinant protein at concentration shown. (K) qPCR analysis showing relative expression of myosin heavy chain isoform genes. Scale bars: 40 μm.