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. 2006 May 1;20(9):1100–1109. doi: 10.1101/gad.1397406

Figure 4.

Figure 4.

Transcription by RNAP II results in an increase in mRNA yield and splicing kinetics. (A) T7 RNA polymerase and T7 DNA template were incubated in nuclear extract for 30 min in the absence (lane 1) or presence (lane 2) of Actinomycin D. (B) T7-DoF DNA template was incubated in nuclear extract in the absence (lane 1,2) or presence (lane 3,4) of T7 RNA polymerase. Actinomycin D was added at 30 min and incubation continued for 0 min (lanes 1,3) or 40 min (lanes 2,4). The corresponding CMV-DoF DNA template was incubated for 30 min; Actinomycin D was added and incubation continued for 0 min (lane 5) or 40 min (lane 6). Total RNA was fractionated on an 8% denaturing polyacrylamide gel. The percent pre-mRNA spliced (mRNA at each time point/pre-mRNA at 30 min) was quantitated by PhosphorImager and is shown in the graph. (C) Same as B except that transcriptions were carried out for 5 min, followed by the addition of Actinomycin D and continued incubation for 15 and 30 min.