Fig. 7
From: Rab32 connects ER stress to mitochondrial defects in multiple sclerosis
Mechanisms of Rab32-mediated neuronal killing. a SH-SY5Y cells were transfected with pIRES2-EGFP, or pIRES2-EGFP expressing Flag-tagged Rab32WT, Rab32Q85L, and Rab32T39N. After 24 h in culture, cells were processed for Cy5-annexin V and stained with DAPI. Scale bars 20 μm. b Western blot analysis showing the expression of Rab32, RIPK1, actin, and tubulin in SH-SY5Y cells transfected with pIRES2-EGFP, or pIRES2-EGFP expressing Flag-tagged Rab32WT, Rab32Q85L, and Rab32T39N. c Quantification of surviving SH-SY5Y cells transfected with pIRES2-EGFP, or pIRES2-EGFP expressing Flag-tagged Rab32WT, Rab32Q85L, and Rab32T39N either under control conditions (black) or treated with autophagy (bafilomycin, red), necroptosis (necrostatin-1, gray), apoptosis inhibitors (zVAD-fmk, blue), as well as a combination of necrostatin-1 and zVAD-fmk (green). After 24 h in culture, the cells were fixed, and surviving neuronal cells were quantified. In each plasmid group, each treatment was compared to the untreated control (No-inhibitor). n = 3