Fig 2.

RNA polymerase II positioning in SAMD4A, EXT1, and ZFPM2 during a transcription cycle. HUVECs were treated with TNF-α for 0 to 85 min, and ChIP followed by qPCR were applied. In some cases, the transcriptional inhibitor, DRB (50 μM), was added 25 min before harvesting. Diagrams on the left illustrate regions targeted by primer pairs used and their distance (in kbp) from the TSS. Bound RNA polymerase was detected using an antibody recognizing the largest subunit of the enzyme. The results (percent enrichments relative to the input ± the standard deviation [SD]; n = 6) are superimposed on data obtained previously using microarrays (44). Peak enrichments at different times coincide with the traveling (yellow) and standing waves (green). *, DRB significantly reduces enrichments seen (P < 0.01; unpaired two-tailed Student t test).