Figure 5

Effect of HIF-1α overexpression on apoptosis in HK-2 subjected to 16 h of hypoxia. (A) HIF-1α protein expression in HK-2 cells transfected with either the empty vector or pcDNA3-HA-Hif1α and subjected to hypoxia or normoxia. Immunoblotting of the same lysates with Hif-1β to monitor protein loading is also shown. (B) Morphological analysis of apoptosis with Hoechst 33258. *P < 0.05 vs hypoxia in pcDNA3. pEGFP was co-transfected alongwith the vector and HIF-1 alpha construct to aid in the identification of apoptotic nuclei in EGFP transfected cells. (C) Apoptotic cells as determined by TUNEL staining. *P < 0.001 vs hypoxia after transfection with pcDNA3. Data shown is mean ± SEM of six separate experiments and (D) Caspase 3 activity (O.D. units 405 nm).*P < 0.001 vs. hypoxia with the empty vector pcDNA3. Data in panels B, C and D are means ± SEM of triplicate wells of six separate experiments.