Fig. 3. Aggregate formation of mtSOD1 and expression of both wt and mtSOD1 induce cell death in mutated astrocytes transfected with GFP-mtSOD1. (A) Intracellular aggregates were induced in astrocytes by incubation with transfected GFP-mtSOD1. Aggregate formation was examined by measuring levels of hSOD1-EGFP fluorescence in the cytoplasmic area at 48 h post-transfection. In WT astrocytes transfected with GFP-mtSOD1, increased aggregate was observed. In MT astrocytes transfected with GFP-mtSOD1, expression of GFP-mtSOD1 significantly increased aggregate in astrocytes. Data are expressed as mean values (±standard error). Scale bar=50 µm. (B) Apoptosis of astrocytes was indirectly assessed by evaluating changes in levels of cleaved caspase-3 by western blotting. Expression of cleaved caspase-3 was largely increased in mtSOD1-transfected astrocytes. In contrast, expression of cleaved caspase-3 was not affected in WT astrocytes with transfected wtSOD1. β-actin was used as the loading control. Astrocytes were transfected with wt and mtSOD1, and increased cell death was observed. In MT astrocytes transfected with GFP-mtSOD1, expression of GFP-mtSOD1 significantly increased apoptotic cell death in astrocytes. Data are expressed as mean values (±standard error).
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