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

Figure 4.

Figure 4

Inhibition of TRPC6 downregulation by HYP9 protects astrocytes from ischemic damage. Astrocytes were pre-incubated with 0, 1, 5, 10, 15, 20, and 30 μM HYP9 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 15 μM HYP9 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 < 0.05, **p < 0.01, ****p < 0.0001 vs. CON (HYP9 = 0 μM) group; #p < 0.05, ###p < 0.001, ####p < 0.0001 vs. OGD (HYP9 = 0 μM) group.