FIG. 4.

HIV induction downregulates host gene expression at the integration site. (A) UBXD8 expression in J-Lat clones. RNA from J-Lat E27 and A2 cells was extracted, and expression of the wild-type UBXD8 transcript was measured by RT-qPCR with a pair of oligonucleotides in exons 8 and 9 (19 plus 20) flanking the HIV integration site at intron 8 in J-Lat E27. As a control, expression of uninterrupted intron 8 was measured with the primer pair 5 plus 10. GAPDH expression was measured for normalization. (B) UBXD8 expression upon HIV reactivation. J-Lat E27 and A2 cells were untreated or treated with TNF-α (10 ng/ml) or TSA (400 nM) for 24 h, RNA was extracted, and transcription through UBXD8 exon 8 was measured by RT-qPCR with oligonucleotides 19 and 21 or through intact intron 8 with oligonucleotides 5 and 10. GAPDH expression was measured for normalization. Data are expressed as fold change (FC) in UBXD8/GAPDH expression in treated cells compared to that in untreated cells. (C) Time course response to TNF-α and PMA. J-Lat E27 cells were treated with TNF-α (10 ng/ml) or PMA (10 nM) for the time points indicated in the figure, and transcription through different UBXD8-HIV regions was measured by RT-qPCR with the primer pairs indicated. GAPDH expression was measured for normalization. Data are expressed as fold change (FC) in primer pair/GAPDH expression in treated cells compared to that in untreated cells (time zero). Values represent the mean and range of representative experiments performed in duplicate.