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. 2016 May 26;11(5):e0156451. doi: 10.1371/journal.pone.0156451

Fig 2. E2F and STAT Transcription Factors Have Predicted Binding Sites and can Bind to the α7 Promoter.

Fig 2

(A) Schematic representation of 1000bp regions of the α3 nAChR, α5 nAChR, and α7nAChR gene promoters showing potential E2F binding sites as black ovals. (B) Schematic representation of 1100bp region of nAChR α7 gene promoter showing potential E2F binding sites as black ovals and potential STAT binding sites as green ovals. Arrows represent the position of primers spanning E2F and STAT binding sites used for ChIP assays. Sequence data of overlapping E2F and STAT binding sites is detailed. (C) ChIP assays showed binding of E2F1-5 to the α7 nAChR promoter region -177 through +13 in A549 cells and binding of E2F1-4 in H460 cells. E2F1 and E2F4, and to some extent E2F2 and E2F3 could also bind region -294 through -463 in A549 cells while E2F1-5 could bind this region in H460cells. STAT1 and STAT3 additionally could bind region -294 through -463 in A549 cells, while STAT1 alone could bind in H460 cells. Sonicated DNA was used as input control, and there was no detectable amplification from irrelevant IgG, used as negative control. Quantification of the data is depicted as percent of input, in the corresponding graphs.