THZ1 inhibited differentiation of mouse primary myoblasts. (A) In mouse primary myoblasts, qRT-PCR was performed to analyze the transcription levels of muscle-specific genes including Myog, Myh3, Mylpf and Mef2c.(B) 3xMef-luc and Myog-luc were co-transfected with pEF-RL respectively. 24 h after transfection, gradient concentration of THZ1 (0, 50, 100 nM) was added. 48 h after differentiation induction, luciferase activity was measured. (C) Protein level of myogenin in mouse primary myoblast was analyzed by western blot. (D) Bar graph was gray intensity analysis of figure 4C. Gray values were normalized to β-tubulin. (*P<0.05, **P<0.01, ***P<0.001, n=3, bars represent SD).