Figure 6
From: Heterogeneous susceptibility of circulating SIV isolate capsids to HIV-interacting factors
HIV and circulating SIV capsids vary in their dependence on Nup358/RanBP2 for the optimal infection of human cells. (A) Specific down modulation of Nup358/RanBP2 in HEK-293T cells by targeted siRNA. Whole-cell extracts of HEK-293T cells transfected with siRNA directed against either the luciferase gene (siLuc) or Nup153 (siNup153) or Nup358/RanBP2 (siNup358/RanBP2), were immunoblotted with a rabbit polyclonal antibody directed against Nup358/RanBP2 (upper panel) or a monoclonal antibody directed against lamin B, used as loading control (lower panel). (B) Infection of Nup358/RanBP2-depleted HEK-293T cells as compared to infection of siLuc-treated cells were determined as ratio of eGFP-positive cells as described in Figure 2 and Figure 3 legends using MLV-B, wt and chimeric HIV-1 constructs, HIV-2ROD, SIVmac, and circulating SIV capsid constructs. (C) Restriction activity in CHO cells stably expressing a synthetic TRIM-RanBP2Cyp fusion protein as compared to CHO cells. Cells were infected as described above with virions harboring CA from wt SIVmac and HIV-1, chimeric HIV-1 constructs, HIV-2ROD, and circulating SIV. All results were obtained from at least three independent experiments with each CA. Unpaired two-tailed Student’s t-test was used to assess significance. SEM and P-values < 0.001 (asterisks) are indicated.