FIGURE 3.

Knockdown of Plin4 decreases LD accumulation and alleviates SH-SY5Y cell damage. (A) SH-SY5Y cells were transduced with siRNA against Plin4 or an SCR control. Western blot analysis was performed at 48 h post-transduction to assess for Plin4 suppression. ^∗∗P < 0.01, ^∗∗∗P < 0.001 vs. SRC as determined by one-way ANOVA followed by the Holm–Sidak test. (B) Cells described in (A) followed by CTL (PBS alone) or 200 μM MPP⁺ stimulation for another 24 h. Live cells were stained with BODIPY^493/503 (2 μg/ml) then fixed and counterstaining with DAPI for fluorescence imaging. (C) Oil Red O staining of the cells described in (B). Right: quantitation of LD number and integrated optical density (IOD). (D) Plin4 knockdown and MPP⁺ stimulation-induced cytotoxicity in SH-SY5Y cells were determined by Hoechst 33342 staining. (E) The cells described in (B) were subjected to the LDH release assay after 24 h of treatment. (F) Annexin V/PI staining indicated apoptosis of the cells described in (E). Data in (C,E,F) are shown as mean ± SEM. ^∗P < 0.05, ^∗∗P < 0.01 vs. SCR-CTL, ^&P < 0.05 vs. SCR-MPP⁺ as determined by two-way ANOVA.