Fig. 4.

LEDGIN treatment shifts HIV-1 proviral localization towards the inner nuclear compartment. U2OS WT or LEDGF/p75 depleted cells were infected with a HIV derived vector pHR-CMV-EGFP with or without an I-SceI site (mock) and γH2AX foci quantified per nucleus 48 h post infection after endonuclease digestion. (a) SCIP analysis of proviral DNA corresponding to γH2AX foci (red) in U2OS cells. Bar diagram in the right panel depicts the number of proviruses (H2Aγ foci) detected under each condition. Error bars represent standard deviations from at least three experiments. (b) 3D nuclear localization of HIV-1 provirus relative to the nuclear rim in LEDGF/p75 KD (empty bars) or WT U2OS cells (grey bars) (p < 0.001, Kolmogorov–Smirnov test) (n = 1000). The right panel depicts the cumulative frequency for the distance relative to the nuclear rim. (c) Cumulative frequency of the 3D nuclear localization of HIV-1 provirus relative to the nuclear rim in U2OS cells treated (grey) with or without (black) 3 μM of LEDGIN CX05045 (p < 0.001, Kolmogorov–Smirnov test) (n = 650). (d) Cumulative frequency of the 3D nuclear localization of HIV-1 provirus relative to the nuclear rim in U2OS cells infected with the HIV-1 INA128T/E170G and treated with (grey) or without LEDGINs (black, CX05045, 3 μM, 4 × IC₅₀) (p > 0.05, Kolmogorov–Smirnov test) (n = 650). Number of experiments is indicated in each plot (> 100 cells counted/experiment).