Fig. 3

MOV10 prevents A3G from Vif-induced degradation by decreasing the ubiquitination of A3G. a Human 293T cells were transfected with pcDNA3.1-A3G-HA (0.8 μg), pcDNA3.1-Vif-HA (0.5 μg), and pcDNA3.1-MOV10-FLAG (1.5 μg) and then treated with MG132 (4 μM) for 16 h. Lysed cells were collected at 48 h and detected by western blotting with anti-HA, anti-FLAG, and anti-GAPDH antibodies. b 293T cells were transfected with pcDNA3.1-A3G-HA (2 μg), pcDNA3.1-Vif-FLAG (1.25 μg), pcDNA3.1-MOV10-FLAG (2.5 μg), and pcDNA3.1-Ub-FLAG (3 μg). Cells were treated with MG132 (4 μM) for 16 h and analyzed by co-immunoprecipitation with anti-HA agarose beads. And then, samples were detected by western-blotting using anti-HA, anti-FLAG, and anti-GAPDH. Values in a represent percentages of A3G normalized against GAPDH and compared with control. The bar graphs in a represent the average expression of A3G with different treatment and relative to the A3G-only reaction control (set to 100%). Data in a represent mean ± SD from three independent experiments. Empty vector pcDNA3.1 was used to equalize DNA amounts in each transfection. Data in a and b are representative of at least three independent experiments