Fig. 6

Evidence of enhanced proinflammatory glial cell phenotype in the striatum of Nlrp3^L351P animals. a Striatal tissue sections from 18-month-old animals were probed with anti-IBA1 antibodies followed by BOND polymer detection. Microglial activation can be visualized in animals harboring the Nlrp3^L351P allele (bottom panel), compared to their WT littermates (top panel). Scale bars represent 20 μM. There are significantly more activated microglia in tissues from animals expressing Nlrp3^L351P compared with tissues from Nlrp3^WT animals (* p = 0.0164, t test). Error bars represent s.e.m. b Adjacent striatal sections were stained with anti-GFAP antibodies. GFAP-immunoreactive astrocytes in Nlrp3^L351P animals have an increased activation phenotype (bottom panel), compared to Nlrp3^WT animals (top panel). Scale bars represent 20 μM. Quantitation revealed significantly more total astrocytes (* p = 0.0217, t test) as well as significantly more activated astrocytes (*** p = 0.0005, t test) in striatal tissues from Nlrp3^L351P animals compared to controls. Error bars represent s.e.m. c Expression of A1 astrocyte markers, Ggta1, H2-D1, C3, were assessed in 18-month-old animals. RNA was isolated from striatal tissues, reversed transcribed, and mRNA was measured by real-time PCR. Ggta1, H2-D1, and C3 levels were significantly elevated in Nlrp3^L351P mice compared to Nlrp3^WT animals (**** p < 0.0001, **** p < 0.0001, ** p = 0.009 respectively, one-way ANOVA). Error bars represent s.e.m.