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. 2014 Jul 1;34(7):526–536. doi: 10.1089/jir.2013.0077

FIG. 6.

FIG. 6.

OM85-BV induced activation and translocation of NF-κB via TLR4 and TLR2 signaling pathway. (A) The activation of P65 and IκB in response to OM85-BV was examined by western blot. n ≥3, *P<0.05 compared with 0 h. (B) Western blot was used to detect the cytoplasmic protein and nucleoprotein P65 in RAW264.7 cells. n ≥3, *P<0.05 compared with 0 h. (C) RAW264.7 cells were pretreated with 5 μM Bay11-7082 for 2 h before 100 μg/mL OM85-BV treatment. RT-PCR was used to analyze IL-1β, IL-6, and TNF-α mRNA expression. ELISA was performed to measure the levels of cytokines in supernatants. n ≥3, *P<0.05. (D) Cells were transfected with TLR4- or TLR2-siRNA and after 48 h they were stimulated with 100 μg/mL OM85-BV for 6 h. The level of P65 and IκB phosphorylation was discovered by western blot. n ≥3, *P<0.05, #P<0.05, and ☆P<0.05. Cells were transfected with MyD88-siRNA or TRAM-siRNA, respectively, for 48 h, and then they were stimulated with 100 μg/mL OM85-BV for 6 h. Western blot was used to detect the level of P65 and IκB phosphorylation. n ≥3, *P<0.05, #P<0.05, and ☆P<0.05. NF-κB, nuclear factor-kappa B.