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. 2014 Feb 15;28(4):342–356. doi: 10.1101/gad.231274.113

Figure 3.

Figure 3.

(A) Diagram of βWT and βpAm genes integrated into HEK293 cells. The tetracycline-inducible CMV promoter (arrow), exons (black boxes), and pA site are indicated. Regions assayed by ChIP are underlined. (B) ChIP across βWT and βpAm to detect the relative ratio of Ser2p as compared with the Pol II (N20) signal. Note that the signal differential between Ex1 and 3′B is significant (P < 0.05) for βWT but not for βpAm. (C) 3′ RACE detection of PCPA of NR3C1 pre-mRNA in total RNA from control and U1 AMO-treated cells. The top panel shows the PCPA product, and the bottom panel shows product from the intronless TAF7 gene that acts as a loading control. The diagram shows the NR3C1 gene and a zoomed region showing primer pairs to detect PCPA within intron 2. (D) ChIP across the 5′ portion of the NR3C1 gene to detect the distribution and level of Pol II, Ser5p, Ser7p, and Ser2p in control or U1 AMO-treated cells. The diagram shows the 5′ portion of the NR3C1 gene, and amplicons are indicated and underlined. For each antibody, values are normalized to the maximal value obtained in control AMO-treated cells. All error bars represent standard deviation from at least three biological replicates.