Figure 6. JQ1(+) treatment increases the MCV replication.
A. JQ1(+) releases Brd4 from chromatin. C33A cells were treated with 300 nM JQ1(+), JQ1(−) or DMSO for 24 h. Cells were fixed and stained with Brd4 antibody and DAPI. B. JQ1(+) treatment increases the MCV replication. C33A cells were transfected with pT+Ori and split into 3 dishes. At 24 h p.t., cells were treated with 300 nM JQ1(+), JQ1(−) or DMSO. Total DNA was extracted at 48 h p.t., and 20 µg of XhoI/DpnI-digested DNA was loaded in each lane for Southern blotting to detect the replicated plasmid. 1.5 µg total DNA samples that have been digested with XhoI but not with DpnI were also analyzed in Southern-blotting to show equal loading of DNA. NTC, no transfection control. All reactions were performed in triplicates. C. JQ1(+) treatment does not affect cell cycle. C33A cells were cultured for 12 h and treated with 300 nM JQ1(+), JQ1(−) or DMSO. Cells were cultured for another 24 h and fixed for FACS cell cycle analysis.