Fig. 1

MOV10 counteracts Vif-mediated degradation of A3G. a, b MOV10 overexpression inhibits Vif-mediated A3G degradation. 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) (a) or different amounts of pcDNA3.1-MOV10-FLAG (from 0.5 to 2 μg) (b). Then, cells were collected and lysed at 48 h, and analyzed by western blotting with anti-HA, anti-FLAG, and anti-GAPDH antibodies. c, d The effect of MOV10 depletion on Vif-mediated degradation of A3G. Cells were transfected with pcDNA3.1-A3G-HA (0.8 μg), pcDNA3.1-Vif-HA (0.5 μg), MOV10-specific siRNA (50 nM) (c) and/or siRNA-resistant MOV10 construct (rMOV10-FLAG) (0.4 μg) (d). After 48 h, cells were collected and analyzed by western blotting assay with anti-HA, anti-FLAG, anti-MOV10 and anti-GAPDH antibodies. Empty vector pcDNA3.1 was used in each transfection to normalize DNA amounts. Values in a–d represent percentages of A3G or MOV10 normalized against GAPDH and compared with control. The bar graphs represent the average expression of A3G with different treatment and relative to the A3G-only reaction control (set to 100%). All the data represent mean ± SD from three independent experiments. Statistical significance was determined using t test: *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001