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. 2014 Apr;88(8):4275–4290. doi: 10.1128/JVI.03287-13

FIG 8.

FIG 8

Role of viral envelope PtdSer in TIM-1- and TIM-4-mediated lentiviral transduction. (A) Blocking of TIM-1-mediated 2.2 1L1L pseudotype transduction by ANX V or D89E. The 2.2 1L1L pseudotype (50 ng p24/ml) was incubated with ANX V or D89E for 1 h prior to transduction at the concentrations indicated in the figure. The vectors were then used for transduction of 293T and TIM-1 293T cells. (B) Blocking of TIM-4-mediated gp64 pseudotype transduction by ANX V or D89E. The gp64 pseudotype (25 ng p24/ml) was incubated with ANX V or D89E for 1 h prior to transduction at the concentrations indicated in the figure. The vectors were then used for the transduction of 293T and TIM-4 293T cells. (C) Blocking of TIM-1-mediated 2.2 1L1L pseudotype transduction by liposomes. TIM-1 293T cells were incubated for 1 h with the liposomes indicated in the figure. The cells were then transduced with the 2.2 1L1L pseudotype (50 ng p24/ml) for 2 h in the presence of the same liposomes. (D) Blocking of TIM-4-mediated gp64 pseudotype transduction by liposomes. TIM-4 293T cells were incubated for 1 h with the liposomes indicated in the figure. The cells were then transduced with the gp64 pseudotype (25 ng p24/ml) for 2 h in the presence of the same liposomes. (E) Blocking by D89E of HLA class I-mediated transduction of 293T cells with the 2.2 pseudotype. The 2.2 pseudotype (100 ng p24/ml) was conjugated with an anti-HLA class I antibody (Ab), followed by 1 h of incubation with D89E at the concentrations indicated in the figure. The vectors were then used for transduction of 293T cells. The experiments whose results are presented in panels A, B, and E were repeated twice in singlicate and twice in triplicate, and the results shown are the averages and standard deviations of one representative triplicate experiment. The experiments whose results are presented in panels C and D were repeated twice in triplicate, and the results shown are the averages and standard deviations of one representative triplicate experiment. For panels A to D, significance was calculated by comparing the results of TIM-1- or TIM-4-mediated transduction without blocking reagents to those with blocking reagents using a two-sample Student t test (*, P < 0.05; **, P < 0.01).