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. 2019 Oct 25;10:2406. doi: 10.3389/fmicb.2019.02406

FIGURE 4.

FIGURE 4

Inflammasome activation is not induced in the infected skeletal myocytes. Mouse C2C12 myoblasts or the differentiated myocytes using horse serum was treated with LPS (1 μg/ml) or IFN-γ (100 ng/ml) for 24 to 72 h. (A) The expression of NLRP3, ASC, procaspase-1, and proIL-1β in the cells were analyzed by immunoblotting with anti-NLRP3, anti-caspase-1 (p10) or anti-IL-1β antibody, respectively. (B,D) The differentiated myocytes were treated with LPS or IFN-γ for 48 h, and infected with WT C. perfringens (strain 13). The activation of caspase-1 was analyzed by immunoblotting (B). (C) Culture supernatants of the infected myocytes were analyzed for LDH release at 1 to 3 hpi. (D) The differentiated myocytes were infected with WT C. perfringens, and/or any of its isogenic mutants, Δplc, Δpfo and ΔplcΔpfo, and analyzed the LDH release from infected cells. Data are presented as the means ± SD of triplicate samples.