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. 2021 May 24;11:10790. doi: 10.1038/s41598-021-89039-0

Figure 6.

Figure 6

ICA equipotently induced myotube hypertrophy. Differentiated C2C12 cells were exposed to various concentrations of EE (100–200 μg/ml), ICA (1–5 μM), or IGF-1 (20 ng/ml) in serum-free medium for 1, 2, 24, or 48 h as indicated. (A) Representative images of bright fields (upper panel) and fluorescence MyHC (green) and DAPI (blue) of cells after 48-h incubation (scale bar, 100 μm). (B,C) Myotube diameter was measured by images captured from the light microscopy (N = 75/group). (D) Fusion index analysis indicated the percentage of nuclei in MyHC-positive myotube was increased after EE100, ICA2, and IGF-1 treatment (N = 12 fields/group). (E–H) Representative western blot images of MyHC isoforms and signaling transducers in cells treated with EE (100, 200 μg/ml) or ICA (1, 2, 5 μM) for the times indicated. Their quantitative results are shown at the lower panels (N = 3–4). In (H), picropodophyllin (PPP, 5 μM) or 0.1% DMSO (served as vehicle control) was applied 2 h prior to CTL (basal), ICA (2 μM), or EE (100 μg/ml) treatment (N = 3–5). All data were expressed as mean ± SD. The symbol * stands for p < 0.05 as compared to CTL or CTL-DMSO; the symbol # stands for p < 0.05 as compared to intra-group DMSO.