Figure 6.

TRPC6 protects astrocytes against IR injury in modeled ischemia. Astrocytes were infected with lentiviruses carrying mCherry-WT-TRPC6-null (Vehicle1), FLAG-tagged full-length mCherry-WT-TRPC6 (WT-TRPC6), mCherry-shRNA-TRPC6-null (Vehicle2), or mCherry-shRNA-TRPC6 (sh-TRPC6), and then exposed to control condition (CON) or 3 h OGD and 24 h reperfusion (OGD). (A) Representative scatter plots of astrocytic apoptosis measured by Annexin-V FITC/DAPI flow cytometry in each group. (B) Cell apoptosis rate was analyzed by Annexin-V FITC/DAPI flow cytometry, and the data were shown as mean ± SEM (n = 4). (C) Quantification of OGD/R-mediated cell cytotoxicity by LDH assay. Data were shown as mean ± SEM (n = 4). (D) Immunoblots for Flag, TRPC6, and cleaved caspase-3 of the extracts from control or OGD-treated cortical astrocytes acquiring Vehicle1, WT-TRPC6, Vehicle2, or sh-TRPC6 vectors. β-actin was used as a loading control. (E) Quantification of cleaved caspase-3 protein levels shown in (D). Data were shown as mean ± SEM (n = 3). *p < 0.05, **p < 0.01, ****p < 0.0001 vs. CON + naive group; ^###p < 0.001, ^####p < 0.0001 vs. OGD + naive or OGD + Vehicle1 group.