Figure 7
Overexpression of GCN5 suppresses Bim expression and apoptosis following potassium deprivation or glutamate exposure. (a) DIV 5 CGNs were infected with Ad-GFP or Ad-GCN5 at 100 multiplicity of infection. After 48 h, the infected neurons were subjected to nucleic staining (scale bar=20 μm) to determine infectious efficiency or RT-PCR to determine e2f1, egr-1 and bim mRNA level. (b and c) Forty-eight hours posttransfection, the neurons infected as in panel (a) were subjected to 25K and 5K treatment for 6 h and then to RT-PCR to detect e2f1, egr-1, and bim mRNA level, and WB was performed to detect GCN5, Bim and Caspase 3. L.E.: long time of exposure; S.E.: short time of exposure. (d) Forty-eight hours posttransfection, the neurons infected as in panel (a) were subjected to 25K and 5K treatment for 12 h and the apoptotic rate was determined as in Figures 1b. (e and f) Forty-eight hours posttransfection, the neurons infected as in panel (a) were exposed to glutamate (Glu, 100 μM) in 25K media for 12 h followed by RT-PCR to detect e2f1, egr-1 and bim mRNA level or for 24 h and the apoptotic rate was determined as above. Mean±S.E.M., n=3, *P<0.05. GFP, green fluorescent protein
