FIGURE 6.

3-MA abolishes the protective effect of Plin4 deficiency on primary DA neurons. (A,B) A: SH-SY5Y cells were pretreated with PBS or 3-MA, followed by siRNA transfection 24 h before MPP⁺ was given, and apoptosis was assessed by Annexin V/PI staining and flow cytometry. The data for flow cytometry analysis are presented as a percentage of the cell population by normalizing to MOCK-SCR-CTL samples. (C,D) Cells described in (A) were harvested and proteins were collected for the detection of caspase-3 activity by Western blot. (E,F) E: Primary cultured neurons of the mesencephalon suffered indicated treatments, followed by TH IHC detection; amplifications shown in the right frames for details. F: Representative number of TH⁺ neurons and neurite length compared with SCR-NC. Scale bar as indicated. Data in (B,D,F) are shown as the mean ± SEM (n = 4–6). ^∗P < 0.05, ^∗∗P < 0.01, and ^∗∗∗P < 0.001 vs. SCR-NC-CTL; ^&P < 0.05 vs. SCR-NC-MPP⁺; and ^$P < 0.05,^$$P < 0.01 vs. MOCK-NC-MPP⁺ as analyzed by one-way ANOVA followed by the Holm–Sidak test.