Fig. 5

Effect of CHMP2A on OGD-Rep-induced inflammation and cell death. a Representative Western blot showing total caspase-1, AIM2, ASC, IL-1β, and IL-18 protein levels in PC12 cells infected with CHMP2A-specific shRNA or vector shRNA for 72 h and then subjected to OGD. b Quantification of total caspase-1, AIM2, ASC, IL-1β, and IL-18 protein levels in PC12 cell lysates (n = 4, Student’s t test. *P < 0.05 vs. sh-vector group). c Representative RT-qPCR analyses of AIM2, IL-1β, and IL-18 in PC12 cells transduced with CHMP2A or vector (20 MOI) (n = 3, Student’s t test. *P < 0.05, ***P < 0.001, and ****P < 0.0001 vs. sh-vector group). d Cell viability was determined by the CCK-8 assay after 2 h (***P < 0.001 vs. sh-vector group). e PC12 cells were transfected with increasing doses of CHMP2A constructs (for 72 h) before OGD-Rep and were subjected to OGD or normoxia (Nor) as a control. Representative Western blot showing total caspase-1, AIM2, ASC, IL-1β, and IL-18 protein levels in PC12 cell lysates. f Representative immunofluorescence images of AIM2 (red) and DAPI (blue) double-staining in PC12 cells transfected with Lenti-EGFP vector or Lenti-EGFP-CHMP2A virus before OGD-Rep. Scale bar, 50 μm. g Quantification of total caspase-1, AIM2, ASC, IL-1β, and IL-18 protein levels in PC12 cell lysates. h, i PC12 cells were pretreated with 3-MA (10 μM) or CQ (10 μM) before OGD-Rep. Representative Western blot and quantification of AIM2, IL-1β, and IL-18 in PC12 cell lysates (n = 4, one-way ANOVA. **P < 0.01)