Fig 2: MA attenuates LPS-induced inflammatory effects through inhibition of NLRP3. (a) qRT-PCR was performed to evaluate the mRNA expression of NLRP3, ASC and Caspase-1 in RAW264.7 cells. (b) Western blot analysis was performed to detect the protein levels of NLRP3, ASC, and Caspase-1 in RAW264.7 cells. (c) qRT-PCR was performed to evaluate the mRNA expression of NLRP3 in RAW264.7 cells after siRNA treatment. Cells were first transfected with siRNA for 48 h, followed by treatment with 100mg/L MA for 12h, and finally treated with 1mg/L LPS for 24 h. All subsequent experimental cell treatments in this figure are performed in the same way as this cell treatment. (d) Western blot analysis was performed to detect the protein levels of NLRP3 in RAW264.7 cells after siRNA treatment. (e) qRT-PCR was performed to evaluate the mRNA expression of IL-1β and IL-18 in RAW264.7 cells after siRNA treatment. (f) Western blot was performed to detect the protein levels of IL-1β and IL-18 in RAW264.7 cells after siRNA treatment. (g) ELISA was conducted to detect the levels of IL-1β inflammatory cytokines after siRNA treatment. (h) qRT-PCR was performed to evaluate the mRNA expression of NLRP3, ASC, and Caspase-1 in RAW264.7 cells after siRNA treatment. (i) Western blot was performed to detect the protein levels of NLRP3, ASC, and Caspase-1 in RAW264.7 cells after siRNA treatment. Data are shown as mean ± SEM obtained from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.