Fig. 1. Palmitic acid (PA) inhibits myoblast differentiation while upregulating miR-103-3p expression.
C2C12 cells were pretreated with PA (100 µM) for 24 h before differentiation. (A) The cells were stained with an anti-MyHC antibody (green) and Hoechst 33342 nuclear dye (blue). Scale bar: 50 μm. (B) MyHC-positive areas, differentiation indices, and fusion indices were analyzed as described in the Materials and Methods. (C, D) Representative immunoblots of myogenic factors and TWF1 on day three of differentiation and analyzed densitometry. The immunoblot intensities were normalized to the amount of β-Actin. The values are shown as relative ratios, with the intensity of the normalized vehicle control set to one. (E) The expressions of miR-103-3p (miR-103) were determined after 24 h of PA treatment by qRT-PCR and normalized to the amount of U6. All data are shown as the mean ± SEM (n > 3), where the level of significance is represented as *p < 0.05; ***p < 0.001 vs. controls. MyHC, myosin heavy chain; TWF1, Twinfilin-1; qRT-PCR, real-time qualitative polymerase chain reaction.
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