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. 2003 May 5;100(10):6098–6103. doi: 10.1073/pnas.1031851100

Figure 7.

Figure 7

Expression of DN Akt in SAE cells blocks KGF-induced protective effects. SAE cells were grown in supplied medium and were infected at a multiplicity of infection of 1 with control adenovirus expressing GFP or with adenovirus expressing DN Akt for 24 h. Cells were starved for 3 h in serum-free medium supplemented with 0.5 g/l BSA and 25 mM Hepes. Uninfected cells were left in the absence of KGF, and control virus- and DN Akt-expressing virus-infected cells were treated with KGF (100 ng/ml). All cells shown were exposed for 72 h to hyperoxic conditions (95% O2/5% CO2). (A) Adherent cell number and cell viability were assessed by using trypan blue. (B) PARP cleavage was assessed as indicator of cell apoptosis by Western blotting techniques. Membranes were probed with anti-PARP Ab, stripped, and reprobed with anti-actin Ab to demonstrate equivalent loading of protein in all lanes. Densitometric analysis of the PARP bands was performed, and the ratio of intensity of cleaved band to the total intensity (cleaved + uncleaved bands) × 100 was estimated as percentage of PARP cleavage. Similar results were obtained in two independent experiments.