Fig. 2

dCA induces a repressive chromatin environment on the HIV promoter in latently infected OM-10.1 cells. a Viral production in HIV latently infected OM-10.1 cell treated with ART with or without 100 nM dCA. Cells were split and treated on average every 3 days and capsid production in the supernatant was quantified by p24 ELISA. Data are representative of three independent experiments. b SAHA induced viral production in OM-10.1 cells. After 300 days treatment with ART and ART + dCA, cells (highlighted with “⦿/☒” in panel A) were stimulated with 2.5 µM SAHA for 24 h. Capsid production was quantified via a p24 ELISA. Data are average of 3 independent experiments, and the error bars represent SD of 3 independent. c SAHA induced viral mRNA level in OM-10.1 cell. Cellular sample from panel B also used for RNA extraction, and cDNAs from extracted total RNA were quantified by RT-PCR using primers to the Nef region. Results were normalized as the number of viral mRNA copies per GAPDH mRNA. Viral mRNA generated in the ART control was set to 100%, and the error bars represent the SD of 3 independent experiments. d Distribution of RNAP II on HIV genome DNA. ChIP assay to RNAP II was performed using cells samples from panels B-C. Results are represented as the percentage of input and subtracted the background of the isotype IgG control. Data are average of 3 independent experiments and error bars represent SD of 3 experiments for each primer set. e The chromatin structure of the HIV LTR in latent OM-10.1 cell from sample in panel B and C. f Histone H3 occupancy on HIV promoter DNA determined by ChIP. Results are represented as the percentage of input and subtracted the background of the isotype IgG control. The ORF of GAPDH was used as control. Data are average of 3 independent experiments, and error bars represent the SD of 3 experiments for each primer set. g The level of H3 lysine 27 acetylation (H3K27Ac) on HIV promoter DNA determined by H3K27Ac ChIP. The results are presented as acetylation of H3K27 over total Histone H3 level, after subtraction of the isotype IgG control background. The promoter of RPL-10 was used as the control. Data are average of 3 independent experiments, and error bars represent the SD of 3 experiments for each primer set. h The recruitment of PBAF complex on HIV promoter DNA determined by ChIP to BAF180. Results are presented as percent immunoprecipitated DNA over input, after isotype IgG control background subtraction. Data are average of 3 independent experiments, and error bars represent the SD of 3 experiments for each primer set. The promoter of GAPDH was used as the control. i Recruitment of BAF250 to the HIV promoter by ChIP. Results are presented as percent immunoprecipitated DNA over input, after isotype IgG control background subtraction. Data are average of 3 independent experiments, and error bars represent the SD of 3 experiments for each primer set. The promoter of GAPDH was used as the control. Statistical significance was determined using the unpaired t-test (*P < 0.05, **P < 0.01)