Effect of GPR43 agonist on the expressions of GPR43, muscle genes, and muscle proteins.
Total RNA was extracted and real-time PCR analysis was carried out in order to examine the mRNA expressions of GPR43 in the L6 myotube cells treated with 0.5 mM acetic acid for 30 min or 1.0 μM GPR43 agonist for 15 min or 30 min (A). The expressions of mef2a, mb, ppargc1a, cycs, and sdha genes in the L6 cells treated with 0.5 mM acetic acid or 1.0 μM GPR43 agonist for 30 min were examined by real-time PCR analysis (B). Next, muscle proteins, pAMPK, myoglobin, MEF2A, PGC-1α, pNFATc1, NFATc1, and CaMKKβ in the L6 cells treated with 0.5 mM acetic acid or 1.0 μM GPR43 agonist for 30 min were analyzed by western blotting as described in Materials and Methods (C). Each bar represents the mean ±SE (n = 3–6). Multiple comparisons were analyzed with one-way ANOVA followed by the Tukey-Kramer post hoc test. Statistical differences are shown as *p< 0.05, **p<0.01, compared with non-treated control.