Fig. 3

NLRP3 is degraded through CMA. Brain lysates from mice were used for IP using anti-NLRP3 or anti-LAMP2A antibody. B, C Cell lysates from BV2 cells were used for IP with NLRP3. SB203580 increased the NLRP3/LAMP2A and NLRP3/HSPA8 interaction and showed in C. D, E Cell lysates from BV2 cells were used for IP with anti-Flag antibody. Wild-type NLRP3-Flag but not NLRP3AA-Flag was coimmunoprecipitated with HSPA8. F, G Cell lysates from BV2 cells were immunoblotted to determine the levels of LAMP2A and NLRP3 after starvation. The protein levels of LAMP2A and NLRP3 were statistically analyzed in G. Mean ± SEM, n = 3, *p < 0.05. H, I Cell lysates from BV2 cells were immunoblotted to determine the levels of LAMP2A and NLRP3 after treatment with AR7 and QX77. The protein levels of LAMP2A and NLRP3 were statistically analyzed in L. Mean ± SEM, n = 3, *p < 0.05. J BV2 cells were treated with LAMP2A siRNA. Cell lysates from BV2 cells were immunoblotted to determine the levels of NLRP3, ASC and CASP1 after treatment with SB203580. LAMP2A siRNA effectively blocked SB203580-induced NLRP3 degradation. K, L IHC demonstrating decreased LAMP2A protein levels were rescued by SB203580 in the SNpc of α-synucleinA53T-tg mice. Statistical analysis of the scores of LAMP2A staining is shown in L. *p < 0.05