FIGURE 4.

HIV-1 Vpr interacts with MCM10 on chromatin. HeLa-iMock and HeLa-iFlag-Vpr cells were treated with doxycycline for 20 h. Cells were then harvested and lysed in 0.5% Triton lysis buffer and centrifuged to obtain a clear soluble fraction. Pellet was washed and treated with Benzonase DNase to release the chromatin-associated proteins (chromatin fraction). Half of each fraction was treated with 25 μg/ml ethidium bromide, and all the treated and non-treated fractions were subjected to immunoprecipitation (IP) using anti-FLAG antibody conjugated to agarose beads.