FIG 3.

Induction of Tax expression after unintegrated HIV-1 DNA has been epigenetically silenced reverses silencing and rescues viral gene expression. (A) Tax-inducible CEM-SS cells were infected with WT or IN⁻ HIV-1 and treated with nevirapine at 16 hpi to prevent viral spread; then, treatment with or without Dox was added at 24 hpi to induce Tax expression. Cells were harvested to assay NLuc expression at the indicated time points. n = 3 biological replicates. Error bars show SD. Cells were also harvested for ChIP-qPCR to determine the levels of H3Ac (B) and H3K9me3 (C) on the HIV-1 LTR promoter, the viral gag gene, and the NLuc indicator gene inserted in place of nef in NL-NLuc. ChIP levels are expressed as percentage of input. n = 3 biological replicates. Error bars show SD. ***, P < 0.001, calculated by two-way ANOVA with Tukey’s multiple-comparison test.