Figure 6

HDAC-1 is recruited to the HIV-1 LTR by TAR derived miRNA. 293 cells were transfected with pLTR-Luc and either siGFP or siTAR1. After 48 hours cells were crosslinked with formaldehyde, lysed, sonicated and immunoprecipitated for HDAC-1 or a IgG antibody. A) PCR on DNA from HDAC-1 ChIP, control IgG ChIP and input was performed for either LTR or luciferase (Luc) region. B) TZMbl cells were cultured in the presence of 100 nM TSA for 0 or 7 days (lanes 1 and 2), and then allowed to recover in the absence of TSA withouth (lane 3), with either TAR-WT or siTAR1 transfection (lanes 4 and 5), or transfection with siGFP (lane 6). PCR for LTR was performed on HDAC-1 ChIP DNA (top panel), no antibody control (middle panel) or input DNA (bottom panel). C) Densitometry was performed on the gel shown in panel B. Background signal in the no antibody PCR was subtracted from the HDAC-1 ChIP and results are shown as a percentage of HDAC-1 association with the LTR before TSA treatment.