Figure 3

Inhibition of nSMase improves oxidative capacity of C2C12 myotubes and reduces TG storage. (A) Analysis of PGC1α gene expression in C2C12 myotubes cultured under palmitate condition in presence or absence of nSMase inhibitor (GW4869). β-actin was used as housekeeping gene control. (B) Mitochondrial DNA copy numbers were estimated by quantitative real time PCR using total DNA isolated from myotubes cultured under control or palmitate condition in presence or absence of nSMase inhibitor (GW4869). Mitochondrial DNA copy number were quantified by assessing ND1 (mitochondrial gene) level which was normalized to HPRT (nuclear gene). Cellular ATP levels (C) and TG storage (D) were quantified after culturing C2C12 myotubes under similar conditions as described under ‘Methods’ section. Data are presented as mean + standard deviation. n = 4, ***P < 0.001, **P < 0.01, *P < 0.05, one way ANOVA with Newman-Keuls post test was performed for statistical analyses.