Figure 4.

E2 reduces NOX2-containing NADPH oxidase complex formation in hippocampal CA1 after cerebral ischemia. A–C, Protein samples from the CA1 region at 3 h reperfusion were immunoprecipitated with anti-p47phox (P47) antibody and then separately Western blotted (WB) with anti-NOX2, p67phox (P67), or P47 antibody (A). In reciprocal Co-IP experiments, samples were subjected to IP with anti-NOX2, p67, or nonspecific IgG (control; data not shown), and the immunocomplexes were probed for the presence of p47, p67, and NOX2 as indicated. Data were shown as means ± SE from independent animals (n = 4–5) and expressed as fold versus sham. ^†p < 0.05 versus sham and E₂ groups. D, E, Western blot analysis using membrane (M) and total (T) protein samples from 3 h reperfusion CA1 regions with anti-p47phox or anti-p67phox antibody. Blots were then stripped and reprobed with anti-cadherin or anti-actin antibody to show loading control. Data were expressed as fold versus sham from four or five rats. *p < 0.05 versus Pla group.