Figure 2

IID-IN mutants are defective for multi-round and single-round replication: A. Fluorescence microscopic images of CEM-GFP cell cultures infected with HIV-1_NL4-3 carrying WT IN or IID-IN mutants in a multiday infection. CEM-GFP cells were infected with normalized amounts of HIV-1_NL4-3 (25 ng p24) WT or IID-IN mutants. Cultures were imaged for expression of LTR-GFP every 48 hr over a 16-day course of infection. B. Measurement of virus particle release in the culture supernatant by p24 ELISA. Culture supernatants were collected from CEM-GFP cells infected with normalized amounts of HIV-1_NL4-3 virus harboring WT or IID-IN mutant IN, every 48 hr and measured for p24 levels. Green = IID-IN mutants partially defective for INI1 binding; Red = IID-IN mutants highly defective for INI1 binding. Data is representative of two independent experiments. C. Infectivity of HIV-Luc virus harboring WT IN or IID-IN mutations. Graph illustrates luciferase activity induced by infection of HeLa cells with HIV-Luc WT and IID-IN mutants in a single round of infection. HeLa cells were infected with 5 ng p24 of WT or IID-IN mutant HIV-Luc. The infectivity was quantified by measuring luciferase activity 24 hours post-infection. Data represents average of three independent experiments (Mean +/- SEM).