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. 2021 Feb 2;8:594283. doi: 10.3389/fcell.2020.594283

Figure 5.

Figure 5

TRPC antagonist SKF aggravates OGD/R induced astrocytes insults. Astrocytes were pre-incubated with 0, 0.05, 0.5, 5, 10, 20, 30, and 40 μM SKF for 12 h, and then subjected 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/PI flow cytometry in each group. (B) Cell apoptosis rate was analyzed by Annexin-V FITC/PI 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 TRPC6 and cleaved caspase-3 of the extracts from CON or OGD-treated cortical astrocytes acquiring 30 μM SKF or not. β-actin was used as a loading control. (E,F) Quantifications of cleaved caspase-3 and TRPC6 protein levels shown in (D). Data were shown as mean ± SEM (n = 3). **p < 0.01, ***p < 0.001, ****p < 0.0001 vs. CON (SKF = 0 μM) group; #p < 0.05, ##p < 0.01, ####p < 0.0001 vs. OGD (SKF = 0 μM) group.