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. 2020 Nov 6;8:565826. doi: 10.3389/fcell.2020.565826

FIGURE 1.

FIGURE 1

Increased lumican production during myoblast differentiation. (A) Mouse C2C12 myoblasts (MBs) were differentiated into myotubes (MTs) after treatment with 2% horse serum for 3 days. Lumican expression in lysates and conditioned media (CM) of MBs and MTs was determined by western blot analysis. (B) Immunocytochemistry images of lumican (green) and myosin heavy chain (MyHC; red) in MBs and MTs. (C) Quantitative RT-PCR analysis to determine lumican (Lum) and myogenin (Myog) expressions in C2C12 cells with or without treatment with 2% horse serum for the indicated days. (D) Western blot and quantitative RT-PCR analyses of lumican after infection with lumican shRNA CM for 6 h in C2C12 cells and differentiation into MTs after treatment with 2% horse serum for 3 days. (E) CCK-8 assay to assess the viability of preosteoblast MC3T3-E1 cells after exposure to 50% CM collected from MTs with or without lumican deletion. The 10 nM lumican was added in lumican silenced CM for the rescue experiment. Scale bar, 200 μm (B). Data are presented as mean ± SEM. *P < 0.05 vs. control; #P < 0.05 vs. myotube CM-treated group.