Fig. 2

ATE1 knockdown in HIV-1 producer cells decreases the infectivity of progeny viruses. A Confirmation of ATE1 knockdown efficiency. CEM/LAV-1 cells were transfected with control or ATE1 specific siRNA. ATE1 expression levels were confirmed by western blotting at 48 h post-transfection. B Effect of ATE1 knockdown in viral producer cells on the packaging of viral proteins in progeny viruses. The purified viruses were subjected to western immunoblotting. Each viral protein was detected by HIV-1-positive plasma. The arrow indicates the positive band of ATE1. C Effect of ATE1 knockdown in viral producer cells on viral production. Viral production levels were monitored by p24 ELISA of culture supernatant of transfected cells. The value in the control experiment was set as 100 %. D Infectivity of viruses produced from control or ATE1 specific siRNA-treated cells. TZM-bl cells were incubated with an equal amount (1 ng of p24 antigen) of each virus. The infectivity was examined based on the luciferase activity in the lysates of each virus-infected TZM-bl cell sample. The value in the control experiment was set as 100%. E Amount of de novo synthesized R/U5 cDNA products of viruses produced from control or ATE1 specific siRNA-treated cells in TZM-bl cells. TZM-bl cells were incubated with an equal amount (10 ng of p24 antigen) of each virus. F Amount of de novo synthesized R/gag cDNA products of viruses produced from control or ATE1 specific siRNA-treated cells in TZM-bl cells. TZM-bl cells were incubated with an equal amount (10 ng of p24 antigen) of each virus. G Amount of de novo synthesized R/U5 cDNA products of viruses produced from control or ATE1 specific siRNA-treated cells in PBMCs. PBMCs were incubated with an equal amount (10 ng of p24 antigen) of each virus. H Amount of de novo synthesized R/gag cDNA products of viruses produced from control or ATE1 specific siRNA-treated cells in PBMCs. PBMCs were incubated with an equal amount (10 ng of p24 antigen) of each virus. The significance of difference (Student’s t-test) is indicated as follows: **p < 0.01; n.s. not significant. The mean values of at least three independent experiments are shown. The error bars denote the standard deviation