Figure 6.
Nascent RNA within Pol II Complex at TES
(A) Meta-analysis of mNET-seq with indicated Pol II antibodies over TES regions (−0.5 k∼+7kb) from siLuc (dark gray) and siCPSF73 (red) treated HeLa cells (left) is shown. Also shown are RTIs of mNET-seq following CPSF73 knockdown (right). GB signals were divided by signals in a 2 kb region from TES (TES+2k) for RTI (see Extended Experimental Procedures). Dashed line is median of siLuc. (∗∗) p value < 8.52 × 10−11, and (∗∗∗) p value < 2.17 × 10−35 by two-sided Mann-Whitney test.
(B) Meta-analysis of mNET-seq/S2P following termination factor knockdown over TES regions (top). siLuc (dark gray), siCstF64+siCstF64t (blue), and siXrn2 (green). RTI of mNET-seq following indicated knockdown (bottom) is shown. (∗∗) p value < 1.94 × 10−15 by two-sided Mann-Whitney test; ns indicates no difference between samples (p value = 0.9894 by two-sided Mann-Whitney test).
(C) Meta-profiles of ChrRNA-seq following indicated knockdown over TES. siLuc (dark gray), siCPSF73 (red), siCstF64+siCstF64t (blue), and siXrn2 (green).
(D) Model correlating Pol II pausing and PAS-dependent transcription termination at TES. RNA cleavage (scissors) by CPA complex (red circle) at PAS (orange triangle). Pol II elongation speed over 3′ flank region is regulated by PAS recognition on average over a 3 kb region from TES.
For (A)–(C), line and shading represent mean ± SEM for each bin.