Fig. 6.

Estrogen signaling affects on intrinsic immunological behaviors of muscle cells. a Phase contrast microscopical observation demonstrated C₂C₁₂ cells survived and differentiated (2% horse serum) well in IFN-γ-, E₂-, or 4-OHT-added medium. b Western blots analysis showed protein level changes of H-2K^b, H2-Ea, TLR3, and ERβ in HS-differentiated C₂C₁₂ myotubes, in the presence of IFN-γ, E₂, or 4-OHT. The relative band intensities from western blots experiments were normalized to the level of GAPDH and analyzed with ImageJ software. c mRNA levels of IL-1, IL-6, MCP-1, and IL-10 were quantified by qRT-PCR in C₂C₁₂ myotubes, in the presence of IFN-γ, E₂, or 4-OHT. All data are presented as mean ± SD (n = 3). One-way ANOVA was used for multiple comparisons. (*p < 0.05 and **p < 0.01). HS horse serum. Bar = 50 μm