HIV-1 Capsid-Cyclophilin Interactions Determine Nuclear Import Pathway, Integration Targeting and Replication Efficiency (original) (raw)
Figure 6
CypA inhibition forces HIV-1 into a Nup358/Nup153 independent nuclear entry pathway.
A, Titers of VSV-G pseudotyped NL4.3 HIV-1 GFP vectors bearing wild type or mutant CA on HeLa control cells (SC), or Nup358 or TRN-SR2 depleted cells in the presence or absence of 8 µM Cs (mean and SD, n = 3). Relative changes from titer on control cells are shown above the bars. B, HeLa cells expressing scrambled control (SC), or Nup358 specific shRNA were transiently transduced with either MLV empty vector control or vector expressing CypA specific shRNA and subsequently infected with wild type NL4.3 HIV-1 GFP vector or CA mutant P90A or O-group MVP5180 GFP vector. Infection was measured 48 hours later by FACS to enumerate infected cells. Fold changes to control cells are shown. A parallel western blot for experiments in B detecting CypA and β-Actin as loading control. C, HeLa control cells (SC) or transiently Nup153 depleted cells were infected with indicated vectors in the presence or absence of 8 µM Cs (Mean and SD, n = 3). Western blot detecting Nup153 and β-Actin as a loading control. D, MDM from 2 independent donors were infected with 400 pg RT replication competent HIV-1 NL4.3 (Ba-L Env) in the presence of 5 µM Cs or DMSO. Replication was measured over time by counting CA positive cells after staining with a CA specific antibody.