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

FIGURE 2.

FIGURE 2

The inflammasome activation induced by Clostridium perfringens is mediated by the pore-forming toxin, perfringolysin O (PFO). BMDMs were primed with LPS for 2 h, and infected with WT C. perfringens (strain 13) and/or any of its isogenic mutants, Δplc (α-toxin mutant), Δpfo (PFO mutant) and ΔplcΔpfo (double-mutant of α-toxin and PFO). (A) 1.5% (w/v) agar plates containing brain heart infusion (BHI) medium and 5% (v/v) defibrinated sheep blood show different degrees of hemolysis induced by the C. perfringens strains. Bar, 1 cm. (B) Activation of caspase-1, and cleavage of IL-1β or GSDMD in the infected BMDMs was analyzed by immunoblotting with anti-caspase-1 (p20), anti-IL-1β and anti-GSDMD antibody, respectively. (C) IL-1β release from the infected BMDMs into the culture supernatants at 20–60 mpi was analyzed by an ELISA. (D) Culture supernatants from infected BMDMs were analyzed for LDH release at 20–60 mpi. Data are presented as the means ± SD of triplicate samples.