Fig. 4

Effect of CSN3 knockdown on the expression of myogenic markers. a Representative immunoblots of myogenin and MHC expression in C2C12 cell lines from samples run on a single gel. b A histogram showing the quantification of myogenin expression normalized to tubulin. c Quantification of MHC normalized to tubulin. The shNT-control, shCSN3-Med and shCSN3-Low stable cell lines were cultured in growth media until 70–80% confluence (0 day) and switched to differentiation media (DM) from 1 to 9 days. Total protein (18 μg) was analyzed by immunoblot using myogenin and MHC antibodies. All blots were reprobed for tubulin (internal control). UD, undetectable. Values represent a mean of 4 independent experiments ± SEM. *p <0.05, **p <0.01 and ***p <0.001 versus time matched shNT-control, by two-way ANOVA. d Representative immunoblots of CDK6, p21 and p27 nuclear fractions (left) and cytoplasmic fractions (right) from samples run on a single gel. Cells treated with 1 μg/ml lipopolysaccharide (LPS) were used as internal controls to verify immunoreactive bands