FIG 6.

Components of the Polycomb repressive complex occupy the HIV promoter in a latently infected primary T-cell model and validation of EZH2 inhibition by GSK343 in uninfected PBMCs. PRC1 (RING1B and BMI1) and PRC2 (EZH2 and EED) components occupy the HIV-1 promoter in latently infected primary T cells. (A) NL4-3-infected primary T cells at day 4 and day 8 postinfection (40), as described in the legend of Fig. 7A. ChIPs for total histone H3, H3K27me3, p50, H3K27me2, and mock IgG are also shown. Means and standard errors of the means from three independent experiments are displayed. For each chromatin occupancy described, the Student paired t test was done for 3 pairs of assays. *, P < 0.05 for mock IgG-treated versus untreated cells at day 4; •, P < 0.05 for mock IgG-treated versus untreated cells at day 8; NS, not significant (P > 0.05). (B) Representative blot showing a reduction in H3K27me3 in total uninfected PBMCs at 2 μM GSK343 for 96 h. Histone H3 and α-tubulin are shown as loading controls. (V, vehicle control [0.02% DMSO]). (C) ChIP changes in the samples described in panel B. Occupancies of H3K27me3 and control IgG were measured at the host gene promoters SAT2 (heterochromatic gene) and MyoD (euchromatic region gene). Results were compared with those for DMSO-treated cells by using Student paired t tests. Error bars represent standard errors of the means (n = 3). *, P < 0.05.