Fig. 1: A3-mediated restriction of HIV-1 integration.
From: Antiretroviral APOBEC3 cytidine deaminases alter HIV-1 provirus integration site profiles
A Western blot analysis of virus producer cells. HIV-1 pseudotyped virus was produced in 293 T cells by co-transfection of plasmids coding for NL4-3-ΔEnv/ΔVif/eGFP, VSV-G, and either empty pcDNA 3 plasmid (HIV-1 no A3), or each of the A3 expression plasmids. The Control lane is the transfection of the pcDNA in absence of virus. Cell lysates were subjected to SDS-PAGE and Western blot analysis. B CEM-SS cells were infected with the amounts of virus as indicated after normalization to capsid (p24CA) protein, as determined by ELISA. Infection was measured as the percentage of eGFP+ cells by flow cytometry. Data are presented as mean values ± SD. C Integrated provirus in the CEM-SS cells from B was quantified using Alu-based PCR combined with nested qPCR. Data are presented as mean values ± SD. D 293 T cells were transfected for the expression of each of the A3 proteins, HIV-1 (no A3), or pcDNA (cells transfected with pcDNA plasmid). Viral and cell lysates were mixed together and subjected to co-immunoprecipitation using an anti-FLAG antibody and analyzed by western blotting. Data shown are representative of three independent experiments. Source data are provided as a Source Data file.
