Fig. 6: Microglia-specific IRAK-M ablation aggravates NLRP3 inflammasome-related activity and GSDMD-mediated pyroptosis in vitro. | Cell Death & Disease

Fig. 6: Microglia-specific IRAK-M ablation aggravates NLRP3 inflammasome-related activity and GSDMD-mediated pyroptosis in vitro.

From: IRAK-M suppresses the activation of microglial NLRP3 inflammasome and GSDMD-mediated pyroptosis through inhibiting IRAK1 phosphorylation during experimental autoimmune encephalomyelitis

Fig. 6

Stimulation of 100 ng/mL LPS for 3.5 h followed by 2.5 μM nigericin for 1 h was performed to mimic EAE pyroptosis in primary microglia. A Whole-cell lysate was collected for RT-PCR. The mRNA levels of NLRP3, ASC, cleaved caspase-1, cleaved GSDMD, and cleave IL-1β are presented as relative expression compared to WT microglia (MG) (n = 5 per group). B Western blotting of microglial protein levels of NLRP3, ASC, full-length (FL) and cleaved caspase-1, IL-1β, and GSDMD. The corresponding charts show the quantification of target protein expression (n = 4 per group). C Representative immunofluorescent analysis of NLRP3, ASC, caspase-1, IL-1β, and GSDMD in microglia. Scale bar: 20 μm. The percentage of NLRP3+, ASC+, IL-1β+, caspase-1+, and GSDMD+ microglia counted from ~160 microglia of five independent cultures. Unpaired t-tests or the Mann–Whitney U test were applied, and data are shown as the mean ± SEM. ***P < 0.001.

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