Fig. 3

Mutant alleles expressed in striatal tissues, evidence of NLRP3 inflammasome activity at 18 months of age. a mRNA was isolated from striatal tissues, reverse transcribed, amplified, and sequenced to confirm expression of Nlrp3 allele in target tissues. The alanine to valine substitution at aa350 was observed in mice expressing Nlrp3^A350V (C➔ T, red peak), with evidence of the Nlrp3^WT background copy expression (blue peak). The leucine to proline substitution at aa351 was observed in mice expressing Nlrp3^L351P (TT ➔ CC, blue peaks), with evidence of the Nlrp3^WT background copies expressed (red peaks). bIl1b mRNA transcript was reverse transcribed, amplified, and quantified with real-time PCR technologies. Nlrp3^L351P mice expressed significantly more Il1b mRNA in striatal tissues compared to control animals (**** p < 0.0001, one-way ANOVA, error bars represent s.e.m.). c Histologic sections of substantia nigra were probed with anti-ASC antibodies, followed by BOND polymer detection, and imaged with a brightfield microscope. ASC puncta (red) indicative of NLRP3 inflammasome activity were observed in Nlrp3^L351P mice. (60 × images, scale bar represents 10 μM)