FIGURE 6.

p22^phox and Nox4 (but not other Nox proteins) mediate the anti-apoptotic effect of growth factors in pancreatic cancer cells. MIA PaCa-2 and PANC-1 cells were cultured for 48 h in the absence and presence of IGF-I (100 ng/ml), FBS (15%), or the Akt1/2 inhibitor (50 μm). Cells were transfected with p22^phox or control siRNA (A and C); pcDNA-p22^phoxWT or control pcDNA plasmid (B); or Nox3, Nox4, Nox5, or control siRNA (D and E). A, protein levels of p22^phox were measured by immunoblotting; β-actin served as loading control. A–C and E, DNA fragmentation was measured using cell death ELISA. D, Nox3, Nox4, and Nox5 mRNA levels were measured using real time PCR. In all panels, values are means ± S.E. (n = 3). *, p < 0.05 versus cells transfected with control siRNA (or control pcDNA plasmid) and cultured without growth factors. #, p < 0.05 versus cells cultured in the same condition without inhibitor or siRNA transfection.