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. 2014 Aug 18;9(8):e105204. doi: 10.1371/journal.pone.0105204

Figure 3. Transcription complexes at the MMTV promoter are differentially sensitive to RNA Pol II inhibitors.

Figure 3

(A) DRB and ActD inhibit hormone induced Ras transcription in 3108 or 3617 cells. (B) DRB and ActD have differential effects on GFP-AR and GFP-GR mobility at MMTV LTR. After treatment with agonist R1881 or DEX alone or in combination with inhibitors DRB or ActD, GFP-AR (3108 cells) and GFP-GR (3617 cells) bound to the MMTV-LTR array were subjected to semi-quantitative FRAP analysis. The recovery half time values for ActD treatment were not determined (nd) as the fluorescence recovery was not large enough during the experimental time period. Error bars represent means ± standard error. (C) DRB and ActD decrease GFP-AR and GFP-GR occupancy at the MMTV array. 3108 or 3617 cells were treated as in (A). qPCR was used to validate ChIP analysis performed with an anti-GFP antibody using known binding sites at MMTV LTR. Error bars represent means ± standard deviation. (D) DRB and ActD have distinct effects on elongating Pol II at the MMTV array. 3108 or 3617 cells were treated as indicated in (A). qPCR was used to validate ChIP analysis performed with an anti-Pol II-pSer2 antibody using known binding sites at MMTV promoter. Error bars represent means ± standard deviation.