Figure 3.
Stretch plus 30 μm NMDA nitrates procaspase-3. A–C, Whole-cell lysates from cultures exposed to stretch plus 30 μm NMDA were immunoprecipitated using an agarose-bound monoclonal anti-3-nitrotyrosine antibody and run on immunoblots (middle lanes) along with nonimmunoprecipitated samples from staurosporine-treated cultures as positive controls and from untreated cultures as negative controls. A, Anti-caspase-9 antibody. B, Anti-Apaf-1 antibody. C, Anti-caspase-3 (pro- and active/cleaved forms). n = 2 experiments/blot. The dotted line encompasses the lanes containing the immunoprecipitated sample. The untreated and staurosporine controls were not immunoprecipitated with anti-3-NT antibodies as, in the absence of nitration, these proteins cannot be precipitated with the antibody-agarose conjugate.
