Fig. 6: Astrocytic autophagy deficiency prevented the inhibitory effect of PPX on NLRP3 inflammasome activation in vivo and in vitro.

a–d Astrocytes were incubated with lysosome inhibitor CQ (40 μM) for 2 h, and then subjected to treatments as in Fig. 4d–g. Analysis of Casp-1 (a) and IL-1β (b) cleavage was determined by Western blotting, with group data shown in c (cleaved IL-1β) and d (cleaved Casp-1). n = 3, One-way ANOVA followed by Tukey’s post hoc analysis. *P < 0.05 vs. as indicated; ns, not significant. e A simplified schematic diagram of AAV-control or AAV-eGFP-Cre structure. f Representative images showing a predominant localization of AAV-eGFP-Cre to GFAP⁺ but not Iba1⁺ or NeuN⁺ cells in the striatum of AAV-eGFP-Cre injected C57BL/6 mice at 3 weeks. Scale bar: 20 μm for the upper panel, and 10 μm for lower panel. g Atg5 immunoreactive signal (red) was almost undetectable in AAV-eGFP-Cre infected cells. Scale bar: 50 μm. n = 3. h–k Western blots (h) and group data of cleaved Casp-1 (i), cleaved IL-1β (j), and ASC (k) protein levels in the striatum. n = 4, two-way ANOVA. *P < 0.05, **P < 0.01 vs. as indicated; ns, not significant. One-way ANOVA followed by Tukey’s post hoc analysis. l, m Representative images for SNpc (left panel) and striatum (right panel) regions (l) and quantification of TH⁺ neuron in the SNpc (m). The striatum of Atg5^flox/flox mice was injected with AAV-eGFP-Cre 3 weeks before saline or LPS stereotaxic injection, and PPX was intraperitoneally administered 3 days before and 21 days after LPS injection as Fig. 1e. Mean ± SEM. *P < 0.05 vs. Saline group; ns, not significant. Scale bar: 50 μm. Casp-1 caspase-1.