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. 2020 Aug 14;11:1972. doi: 10.3389/fmicb.2020.01972

FIGURE 6.

FIGURE 6

PM0442 promotes cytokine secretion by interacting with TLR2. (A,B) SDS-PAGE (A) and western blot images (B) for expression and purification of rPM0442 protein of P. multocida. Lane 1: The untranslated protein band, Lane 2: the expression of the rPM0442 protein band induced by IPTG, 3: the protein band after purification by the HIS-Ni column. (C) rPM0442 induces secretion of IL-1β, TNF-α, IL-6, and IL-12p40 in peritoneal macrophages. (D) TLR2 and TLR4 expression after rPM0442 protein treatment. (E) TLR2-specific antibodies reduce the production of IL-1β, TNF-α, IL-6, and IL-12p40 in macrophage induced rPM0442 protein. (F) TLR2– /– macrophages have reduced secretion of IL-1β, TNF-α, IL-6, and IL-12p40 in macrophage induced rPM0442 protein. (G) Pull-down experiment to analyze the binding of rPM0442 to TLR2. Panels (A,B,G) were representative of three independent experiments. Panels (C–F) representative of three independent experiments with 8 replicates per group. Panel (C) was determined by one-way ANOVA. Panels (D–F) were analyzed with by unpaired t-test or Mann–Whitney test. The data significance was expressed as means ± SD (*P < 0.05, **P < 0.01, ***P < 0.001).