FIGURE 3

Effect of hepatocyte growth factor (HGF) on the expression of heat-shock protein 70 (Hsp70) and the interaction of Hsp70 with apoptosis-inducing factor (AIF) in the hippocampal cornu ammonis (CA) subregion 1 (CA1) region after transient forebrain ischemia. (A) Proteins (20 g) from nonoperated naïve rats and four-vessel-occluded rats at 36 h of reperfusion without (I/R) or with HGF (I/R+HGF) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subjected to immunoblotting with anti-Hsp70 antibody. The blots were then stripped and reprobed with anti--tubulin antibody. Bands corresponding to Hsp70 or -tubulin were scanned, and the scanned bands of Hsp70 were normalized by -tubulin on the same blot. Results are the mean percentages of the nonoperated naïve controls.e. (B) Proteins (500 g) from nonoperated naïve rats and four-vessel-occluded rats at 36 h of reperfusion without (I/R) or with HGF (I/R+HGF) were immunoprecipitated (IP) with anti-AIF antibody, and the precipitates were then separated by SDS-PAGE and subjected to immunoblotting (IB) with anti-Hsp70 antibodies. The blots were subsequently stripped and reprobed with anti-AIF antibodies. Bands corresponding to AIF or Hsp70 were scanned, and the scanned bands of Hsp70 were normalized by precipitated AIF on the same blot. Results are the mean percentages of the nonoperated naïve controls.e. n=3. ^* P<0.05 versus nonoperated naïve control, analysis of variance (ANOVA) with post hoc Fischer's protected least significant difference.