Fig. 7

NFAT signaling is involved in the regulation of myoblast differentiation/fusion mediated by Ezrin. A RNA-seq analysis of the CaN-NFAT signaling pathway in C2C12 myoblasts with overexpression or knockdown of Ezrin at 6 days. n = 3. B Western blot of NFATc1-c4 proteins in myoblasts treated with Ezrin overexpression or knockdown for 6 days. C–F Quantitative analysis of NFATc1-c4 protein expression was performed 6 days after myoblast differentiation following Ezrin overexpression or knockdown. Three independent experiments were performed, n = 3, *P < 0.05 vs. Ad-Null; ^#P < 0.05 vs. Ad-Null. G Typical image of MyHC, MyoG or MEF2c staining in differentiated C2C12 myoblasts treated with Ad-shEzrin with or without Ad-NFATc1, Ad-NFATc2, Ad-shNFATc3 and Ad-shNFATc4. Red fluorescence indicates MyHC, MyoG or MEF2c; DAPI indicates the nucleus. H–J Quantitative assay for MyHC + myotubes and MEF2c + or MyoG + nuclei numbers in the above images. Three independent experiments were performed, n = 3, *P < 0.05 vs. Ad-Null; ^#P < 0.05 vs. Ad-shEzrin; ^&P < 0.05 vs. Ad-shEzrin; ^$P < 0.05 vs. Ad-shEzrin