SOD2 Mediates Amifostine-Induced Protection against Glutamate in PC12 Cells

Figure 3

SOD2-siRNA abolished SOD2 protein upregulation induced by amifostine in PC12 cells. The PC12 cells were divided into six groups: control: cells cultured in drug-free medium; Ami: cells exposed to 500 μM amifostine for 24 h; Glu: cells treated with 15 mM glutamate for 24 h; Ami + Glu: cells exposed to 500 μM amifostine plus 15 mM glutamate for 24 h; SOD2-siRNA + Ami + Glu: cells incubated with SOD2-siRNA transfection complexes for 24 h and then exposed to 500 μM amifostine plus 15 mM glutamate for 24 h; SC-siRNA + Ami + Glu: cells incubated with scrambled- (SC-) siRNA transfection complexes for 24 h and then exposed to 500 μM amifostine plus 15 mM glutamate for 24 h. (a) SOD2-siRNA was effective in decreasing SOD2 protein expression. (b)-(c) Amifostine exposure increased SOD2 protein expression, and the effect was reversed by SOD2-siRNA. Western blot analysis and immunofluorescence staining were used to assess SOD2 protein expression. Nuclei were counterstained with DAPI (blue). Results are means ± SD, , NS: no significance, and bar = 10 μm.

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(b)

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