Enhanced Hsp70 Expression Protects against Acute Lung Injury by Modulating Apoptotic Pathways
Figure 4
AdHSP prevents nuclear translocation of activated Caspase-3.
A. Representative autoradiograms for pro-caspase 3 and activated (cleaved) Caspase 3. 30 µg of cytosolic (upper panels) and nuclear (lower panel) extracts were subjected to SDS-PAGE. Immunoblotting performed with primary rabbit polyclonal antibody to pro- caspase 3 and secondary goat anti rabbit IgG, primary goat antibody to β-actin and secondary donkey anti goat IgG, primary rabbit polyclonal antibody to active (cleaved) caspase-3 and secondary goat anti rabbit IgG, primary mouse monoclonal antibody to histone (H1) and secondary goat anti mouse IgG.. β-actin and histone serve as loading controls. B. Representative stained fixed tissue section depicting intra-nuclear staining for Caspase 3. Sections obtained from T0 control, 2CLPPBS and 2CLPHSP rats. Tissue isolated 48 hrs after the induction of sepsis. Upper panel: 40× magnifications. Black arrows indicate active caspase 3 stained nuclei. Lower panel: 100× magnification of upper panel. C. Caspase 3 Activity Assay. Graphic representation of relative caspase-3 enzymatic activity (mean +/− standard deviation) * = significantly different from Control and AdHSP+TNF.