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. 2014 Mar 6;53(5):779–790. doi: 10.1016/j.molcel.2014.01.017

Figure 1.

Figure 1

ChIP-qPCR Analysis Shows Accumulation of pSer5 Pol II over Introns in prp5-1 Cells

(A–H) Cells grown to mid-log phase were shifted from 25°C to 37°C. Samples were taken and formaldehyde crosslinked before the shift (25°C) and after 30 min at 37°C. ChIP was performed using antibodies 4F8 (total RNA Pol II), 3E8 (pSer5), or 3E10 (pSer2) as indicated, followed by qPCR analysis. Solid and dashed lines denote 25°C and 37°C, respectively. Positions of amplicons are shown below each graph. ChIP was performed in WT cells using 4F8 (total RNA Pol II) (A), prp5-1 cells using 4F8 (B), WT cells using 4F8 (C), prp5-1 cells using 4F8 (D), WT cells using 3E8 (pSer5) (E), WT cells using 3E10 (pSer2) (F), prp5-1 cells using 3E8 (G), and prp5-1 cells using 3E10 (H), followed by qPCR analysis. (A), (B), and (E)–(H) used DBP2 (intron-containing), and (C) and (D) used FMP27 (intronless). Error bars indicate SEM from two independent experiments, each assayed in duplicate. Figure S1 shows a time course of Pol II ChIP on DBP2 in prp5-1 cells during incubation at 37°C, plus similar analyses with ACT1 and ASC1.