FIGURE 8.

Endogenous SUMO-2/3 is conjugated to endogenous Nrf2 and to endogenous PML. HepG2 cells were treated with As₂O₃ (2 μm in B or 5 μm in A and C) for various times. Whole cell lysates were subjected to Co-IP assay. Nonimmune serum (IgG) was used as control. A–C, endogenous Nrf2 associates with endogenous SUMO-2/3. A, IP, Nrf2; WB, SUMO-2/3. B, input control. IP, Nrf2; WB, Nrf2. C (reverse Co-IP assay): IP, SUMO-2/3; WB, Nrf2. D–F, endogenous PML associates with endogenous (lanes 1-3) as well as with exogenous (lanes 4-6) SUMO-2/3. D, Western blots of PML performed with 30 μg of whole cell lysate protein. Loading control, β-actin. E, input assay. IP, PML; WB, PML. F, IP, PML; WB, SUMO-2/3. The data in A–F are representative of results from three separate experiments.