FIG 2.

At least one HIV-1 LTR NF-κB binding site is required for the rescue of IN⁻ HIV-1 replication by Tax. Clonal CEM-SS T cell lines with Tet-inducible Tax were infected with WT or IN⁻ NL-NLuc with either neither, one, or both of the NF-κB sites mutated. Live cell counts (A), virally encoded NLuc expression (B), total HIV-1 DNA measured by qPCR (C), and HIV-1 RNA expression measured by qRT-PCR (D) were quantified at 1, 2, 3, 5, and 7 days postinfection (dpi). The WT culture was devoid of live cells at 7 dpi. DNA and RNA levels were normalized to those in WT HIV-1-infected cells in the absence of Tax at 1 dpi, which was set to 1. The epigenetic state of the viral LTR promoter was characterized by quantifying the bound NF-κB subunits Rel A and Rel B (E) and the activating H3Ac and repressive H3K9me3 histone modification (F) at 2 dpi using ChIP-qPCR. n = 3 biological replicates. Error bars show SD. ***, P < 0.001, calculated by two-way ANOVA with Tukey’s multiple-comparison test.