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. 2020 Mar 3;7:37. doi: 10.3389/fmolb.2020.00037

FIGURE 4.

FIGURE 4

MCC950 blocks NLRP3 inflammasome activation post-SCI in vivo. (A) We assessed NLRP3 mRNA levels at 7 days post-SCI. (B) We conducted a CoIP assay to investigate NLRP3 inflammasome assembly, including the mutual combination of NLRP3 and ASC and of NLRP3 and pro-Caspase-1 at 7 days post-SCI. Left: WB analysis of NLRP3, ASC, and Caspase-1 (p20) levels in lysates/input. Right: We conducted the CoIP using an anti-NLRP3 antibody and subjected the CoIP products to WB analysis for NLRP3, ASC, and pro-Caspase-1 levels. (C) IF analysis of IL-1β and NLRP3 levels in the SCI and MCC950 groups. Scale bars: 20 μm. Data are reported as the mean ± SEM, n = 8. One-way ANOVA, **p < 0.01, ***p < 0.001, vs. sham. MCC950 (L: 10 mg/kg, H: 50 mg/kg). CoIP, co-immunoprecipitation; WB, western blot; IF, immunofluorescence; SEM, standard error of the mean.