Fig. 4

C3a induces a pro-inflammatory state via NLRP3 inflammasome in macrophage of PAH rats A–B Increased levels of the p-P65/P65, NLRP3, Cleaved-CASP1, Cleaved-IL1β, and IL18 in lung homogenates of the MCT group as compared to the MCT + CP40-KK group. Representative immunoblot A and graphs summarizing results (B; n = 6 per group). Representative Western blots C and quantification of p-P65/P65, NLRP3, Cleaved-CASP1, Cleaved-IL1β, IL18 in RAW264.7 after treatment with C3a (50 nM) for 24 h (D; n = 3 per group). E, F Representative EdU assay determined the proliferative rate in PASMCs under the CON-supernatant or C3a-supernatant condition plus treatment with PBS or IL-1RA for 24 h (n = 3 per group). Representative Western blots G and quantification of PCNA and Cyclin D1 in PASMCs under the CON-supernatant or C3a-supernatant condition plus treatment with PBS or IL-1RA for 24 h (H,I; n = 3 per group). Scale bar: 100 μm E; mean ± SEM; Multiple comparisons made by one-way ANOVA; *P < 0.05, **P < 0.01