We next examined Nrf2 nuclear binding by electrophoretic mobility shift assays in these macrophages, as Nrf2 is the master transcription factor that activates the antioxidant response element (ARE). As shown in Figure 2, panel B, nuclear binding of Nrf2 was also decreased in macrophages isolated from alcohol-fed rats compared to macrophages from control-fed rats (Lane 3 vs. Lane 1). In contrast, zinc treatment in vitro completely restored PU.1 nuclear binding in macrophages from alcohol-fed rats (Lane 4 vs. Lane 3). Interestingly, and as opposed to PU.1 binding in cells from control-fed rats where zinc treatment had no effect, zinc treatment appeared to increase Nrf2 nuclear binding in alveolar macrophages from control-fed rats (Lane 2 vs. Lane 1).
