Extended Data Fig. 3: Individual clonal knockout cell lines for the indicated SMC5/6 complex components exhibit similar increases in gene expression from IN- HIV-1 and similar growth rates.
From: Epigenetic silencing by the SMC5/6 complex mediates HIV-1 latency
(a) 7-day growth curves for uninfected CEM-SS Cas9 cells, or the ∆SMC5 and ∆SLF2 CEM-SS clones, counting total live cells in culture. (b, c) The indicated cell lines were infected with IN + / IN- HIV-1 and the levels of (B) unintegrated 2LTR DNA, and (c) integrated HIV-1 DNA were quantified by qPCR and Alu-qPCR respectively. DNA levels were normalized to IN + HIV-1-infected CEM-SS Cas9 cells at 1dpi, which was set to 1. Data shown represent the average of three biological replicates with SD indicated. (d)The indicated clonal knockout CEM-SS cell lines were infected with IN + or IN- NL-NLucΔEnv and viral NLuc expression then assayed at 2 dpi. NLuc levels are given relative to WT CEM-SS Cas9 cells infected with IN- NL-NLucΔEnv (Control), which was set at 1.0. Average of three biological replicates with SD indicated. Data was analyzed using one-way ANOVA, Fisher’s LSD test (p-values indicated on graph, ****p < 0.0001).
