Fig. 6

FUS occupies the HIV promoter and its depletion enhances the recruitment of SEC and P-TEFb to the viral promoter. a + b FUS occupancies the HIV promoter. ChIP material was isolated from Jurkat (J)-LTR-Luc or J-LTR-Tat-Luc cells, where the LTR promoter either harbors TAR (a), or consists of a LTR-delta TAR (b). Cells were subjected to FUS-IP using FUS antibody. qPCR on IP samples was conducted with a pair of primers located on the HIV promoter and signals were presented as percentage from input. The error bars represent mean ± SD from three independent qPCR reactions. Asterisks indicate levels of statistical significance calculated by two-tailed student T test (p < 0.01). c FUS restricts AFF4 occupancy on the HIV promoter. ChIP material from Jurkat (J)-LTR-Luc cells and J-LTR-Luc-FUS KO, where FUS expression was depleted and also express HA-AFF4 or HA-AFF4-300, was subjected to immune-precipitated with anti-HA-antibody or control antibody followed by qPCR with primers located on the HIV promoter. Data are presented as percentages of the input DNA, and representatives of three independent experiments. The error bars represent mean ± SD from three independent qPCR reactions. Asterisks indicate levels of statistical significance as calculated by two-tailed student T test (**p ≤ 0.01). d FUS modulates Cdk9 occupancy on the HIV promoter. ChIP-qPCR analysis of J-LTR-Luc, or J-LTR-Luc-FUS-KO cells were performed by using anti-Cdk9 IgG. As control, IP on ChIP material was also performed with non-specific IgG. qPCR analysis was performed with the primers located on the HIV promoter (a). Signals present percentage from input DNA, and results are representative of three independent experiments. The error bars represent mean ± SD from three independent qPCR reactions. Asterisks indicate different levels of statistical significance as calculated by two-tailed student T test (**p ≤ 0.01)