Fig. 8. GAPDH is S-nitrosated during H₂O₂-induced apoptosis. (A) Detection of S-nitrosated GAPDH by immunoprecipitation using an anti-CSNO antibody in cell extracts from untreated cells, H₂O₂-treated (1.0, 1.5 and 2.0 mM) cells and cells treated for 200 minutes with 2 mM of the NO donor diethylenetriamine/NO (DETA/NO). (B) Quantification of band intensity from A by densitometry. Band intensities were normalized to the intensity of IgG bands. Data express the GAPDH/IgG fold change in comparison to control (lane 1). ^*P0.05 versus control, t-test, n=3. (C) Immunoprecipitation of S-nitrosated GAPDH with an anti-CSNO antibody from cellular extracts of untreated, H₂O₂-treated (1.5 mM), either in the absence or presence of L-NAME, or chronologically aged cultures (2 and 5 days). (D) Quantification of band intensity from C by densitometry. Band intensities were normalized to the intensity of IgG bands. Data express the GAPDH/IgG fold change in comparison to control (lane 1). ^*P0.05 versus control, t-test, n=3.