Fig. 4

Linc-RAM enhances muscle cell differentiation in a PYGM-dependent manner. A Representative images of immunostaining for MyoG (green) in C2C12 cells subjected to overexpression of Linc-RAM (OE) with simultaneous siRNA knockdown of PYGM, followed by induction of differentiation for 24 h. Scramble RNA served as the siRNA control (Scramble). The empty vector served as the negative control (Ctrl) for overexpression of Linc-RAM. DAPI (pseudo-colored red) served to visualize nuclei. Scale bars, 200 μm. B Relative expression of MyoG in the cells described in (A), as determined by RT–qPCR. C Representative images of immunostaining for MHC (green) in C2C12 cells subjected to overexpression of Linc-RAM (OE) with simultaneous siRNA-mediated knockdown of PYGM, followed by induction of differentiation for 48 h. Scramble RNA served as the siRNA control (Scramble). The empty vector served as the negative control (NC) for overexpression of Linc-RAM. DAPI (pseudo-colored red) served to visualize nuclei. Scale bars, 100 μm. D Numbers of MHC-positive (MHC+) cells per view calculated in the cells described in (C). All images are representatives of three independent experiments. Values presented indicate the means ± SE obtained from three independent experiments, and were compared by two-way ANOVA. *p < 0.05, **p < 0.01. NS: statistically non-significant