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. 2021 Jan 28;11:618992. doi: 10.3389/fphar.2020.618992

FIGURE 8.

FIGURE 8

Kir6.2-deficient A1 astrocytes fail to induce injury of mesencephalic neurons in vitro. (A) Protocol of treatment for (B–I). The mesencephalic primary neurons were incubated with the ACM mixed with neurobasal medium at a ratio of 1:2 for 12 h to conduct the bioassay. (B) Cell viability of mesencephalic primary neurons was assayed using CCK8 kit. (C) Representative immunofluorescent stainings of MAP2 in primary neurons. (D) Representative images of Hoechst-stained nuclei in primary neurons. (E) Flow cytometric analysis of primary neurons stained with AV/PI kit. (F) Quantification of Hoechst-positive cells was analyzed. (G) Quantification of dead cells in the flow cytometric analysis of AV/PI was analyzed. (H) Bcl-2, Bax, TH and DAT in the cell extracts of primary neurons were analyzed by immunoblotting analysis. (I) Densitometric analysis of Bcl-2, Bax, TH and DAT. Data are analyzed using two-way ANOVA. *p < 0.05, **p < 0.01, and ***p < 0.01 vs. corresponding CON-ACM group. # p < 0.05, ## p < 0.001, and ### p < 0.001 vs. WT A1-ACM group. Values are means ± SEM from three independent experiments.