Fig 1. Restriction by exogenous or endogenous TRIM5α is inefficient in HEK293T cells.
(A) HEK293T and THP-1 cells were retrovirally transduced with WT huTRIM5α, R332G-R335G huTRIM5α or with the “empty” vector as indicated. Untransduced cells were eliminated by hygromycin treatment, and the cell populations were then challenged with increasing amounts of the HIV-1NL-GFP vector. The percentage of cells expressing GFP was then determined by FACS.(B) HEK293T Jurkat cells were infected with increasing amounts of N-MLVGFP and B-MLVGFP. The amounts of virus used are expressed as multiplicities of infection (MOI) as calculated in the non-restrictive CRFK cells. The percentage of infected cells was determined by FACS. For N-MLV in Jurkat cells, only the 3 virus doses that yielded detectable infections are shown. (C) HEK293T cells were treated with IFN-α, IFN-β or IFN-ω for 16 h prior to a single-dose infection with N-MLVGFP and B-MLVGFP, designed to yield 10–20% infected cells in the absence of IFN-I. The percentage of infected cells was determined by FACS.