Analysis of 4E-BP2 by two-dimensional gel electrophoresis and changes induced by ischemia and IR stress.

<p>(A) Samples of cerebral cortex from control (SHC and SHC3d) and ischemic animals, without (I15) or with reperfusion (R30 and R3d), were subjected to two-dimensional gel electrophoresis (2-DGE) and western blotting with an anti-4E-BP2 antibody. The antibody-reactive spots were <i>a</i>’, <i>a</i>”, <i>a</i>”’, <i>b</i>’, <i>b</i>”, <i>b</i>”’ and <i>b</i>””. Figures show representative results from six different animals. (B) Identification of the phosphorylation sites for 4E-BP2 spots. R30 sample as in A was subjected to 2-DGE and western blotting for phospho-specific antibodies against 4E-BP2 Thr³⁷/Thr⁴⁶ (p-Thr37/46). The figure shows a representative result from three different animals. Arrows indicate the relative position of the <i>a</i> and <i>b</i> forms of 4E-BP2 in the molecular weight (MW) axis. MW is indicated in kDa.