FIGURE 3.

GFP-Nrf2 co-localizes with endogenous PML protein and wild-type RFP-SUMO. HepG2 cells grown in MEM on coverslips to 50–80% confluence were co-transfected with 2 μg of pEGFP-Nrf2 with 0.5 μg of expression plasmid for RFP-SUMO-1 or RFP-SUMO-2 followed by immunostaining for endogenous PML protein, as described under “Experimental Procedures.” A, RFP-SUMO-1; B, RFP-SUMO-2; inset in panel 5 is an enlargement of the boxed area within panel 5. Panels 7–9 are representative images of cells from parallel experiments showing co-localization as in panel 5. Yellow arrows point to yellow dots representing co-localization of GFP-Nrf2 with RFP-SUMO; white arrows point to white dots representing co-localization of GFP-Nrf2 with RFP-SUMO and endogenous PML. Scale bars, 10 μm.