FIGURE 2.

α-Synuclein does not substantially affect endoplasmic reticulum or Golgi morphology. A, HeLa cells were cotransfected with cDNAs encoding azurite, mitoGFP, and either vector control (−syn) or α-synuclein (+syn). 48 h after transfection, cells were fixed, and the endoplasmic reticulum was identified by staining for the KDEL receptor. α-Synuclein has no effect on the morphology of the endoplasmic reticulum. Scale bar indicates 10 μm. B, HeLa cells stably expressing mitoGFP were cotransfected with cDNAs encoding azurite and either vector control (−syn) or α-synuclein (+syn). Control transfected cells were also treated with 5 mg/ml brefeldin A (BFA) for 60 min before imaging and then imaged again 4 h after wash out. 48 h after transfection, cells were fixed and stained for the Golgi matrix protein GM130. Random cells were classified blind to transfection in terms of mitochondrial and Golgi morphology. α-Synuclein produces mitochondrial fragmentation (p < 0.0001), whereas control, BFA, and BFA following wash do not. In contrast, BFA causes dramatic fragmentation of the Golgi complex far greater than that by synuclein (p < 0.0001), and this does not affect mitochondrial morphology. n = 24–111 cells per group from four independent transfections.