Fig. 4: Effect of basal ganglia derived ECs on HIV reactivation in microglia (huglia, HC69) cells.

A) Representative microscopic images of internalized DIR-labeled ECs by huglia cells (top) and the level of internalized ECs (bars, bottom). Black squares represent enraged images linked with broken lines. B) The levels of latent HIV LTR promoter transactivation (GFP expression) by control molecules, including PBS, dexamethasone (DEX) and TNFα). C) Concentration (1 µg, 2 µg, 3 µg, 4 µg, 5 µg in 100 µL)-dependent assessment of the potency of ECs from different sources in transactivation of latent HIV LTR promoter at 96 h after treatment. D) 2-way ANOVA (Šídák’s Multiple Comparison, simple effects within rows) analysis for the data in panel “C”. E) Quantification of latent HIV LTR promoter transactivation (GFP expression, green) at different times (days 1, 2, 3, 4) after treatment with VEH, VEH | SIV, THC | SIV ECs and other compounds (PBS, DEX, TNFα). F) Representative microscopic images of huglia cells on day 4 after treatment with VEH, VEH | SIV, THC | SIV ECs and other compounds (PBS, DEX, TNFα) showing latent HIV LTR promoter transactivation (GFP expression, green). G) Representative microscopic images of hμglia cells showing THC | SIV ECs suppressing TNFα-mediated HIV LTR promoter reactivation. H) The levels of HIV reactivation (GFP expression) by TNFα and suppression by THC | SIV ECs. I) Representative microscopic images of hμglia cells showing VEH | SIV ECs reversing DEX-mediated inhibition of HIV LTR promoter reactivation. J) The levels of inhibition of HIV reactivation (GFP expression) by DEX and reactivation by VEH | SIV ECs. All experiments were repeated three times. Statistical differences were assessed by ordinary 2-way ANOVA with Tukey’s multiple comparison test as presented on Tables 4 – 6. ns = non-significant. Scale bars (horizontal orange lines) = 200 µm.