FIGURE 2.

NRF2 activity is increased in a dose-dependent manner when Ref-1 redox activity is inhibited via E3330. Pancreatic cancer cells, PaCa-2 (A) and Pa02C (B), were assayed for NRF2 activity via luciferase reporter assay. At 24 h post-transfection of luciferase and firefly constructs, PDAC cells were treated with vehicle control (DMSO), E3330, or negative control compound, RN7–58, for 24 h and then the luciferase assay was conducted. Doses were based on survival data from previously published data (4). C, quantitation of HMOX-1 expression by QPCR using PaCa-2 cells transfected with Ref-1 siRNA#1 or scrambled control and then treated with E3330 at 50 μm for 24 h; and D, immunoblot for Ref-1 for the samples in C with Actin as loading control. *, p < 0.05, comparing DMSO control to drug-treated samples; n = 3–4.