Localization of 1-LTRc formation. (A) Left panel, HeLa cells were infected cells with NLENG1-ES-IRES D116N and fractionated into a cytoplasmic and nuclear fraction 24 h post-infection. 2-LTRc (black columns), 1-LTRc (grey columns) and DNAL (white columns) were quantified in each fraction. Right panel, quantification of ß-globin gene and mitochondrial gene in the nuclear and cytoplasmic compartment. (B) HeLa cells were infected (WT + 500 nM RAL; black columns) in the presence of either 100 μM NLS-IN-Pen (dark grey columns) or 100 μM SV40-NLS-Pen (grey columns). Percentages of 2-LTRc (left panel) and 1-LTRc (right panel) at two times post-infection are shown. (C) HeLa cells were infected either with pNL4.3 virus (WT; black columns) or the CTS/cPPT mutant virus (grey columns). Ratios of 2-LTRc and 1-LTRc (the amount of 2-LTRc or 1-LTRc in the mutant condition divided by that for the WT at 24 h post-infection) are shown. Each value corresponds to an average of five to six independent experiments and confidence intervals analysis are shown for a p value <0.05.