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. 2013 Aug 13;8(8):e72496. doi: 10.1371/journal.pone.0072496

Figure 5. The efficiency of transcription reinitiation depends on the length of the transcripts.

Figure 5

(A) In vitro transcription was performed using the pfMC2AT templates harboring a G-free cassette that ranges from 350 to 50 nt in length. The numbers above each lane indicate the length of the G-free cassette. Mediator and hnRNP R were added as indicated in the figure. A representative data of three experiments is shown. (B) Quantification of the first-round transcripts and the second-round transcripts in the presence of Mediator alone (“Mediator ONLY”) or both Mediator and hnRNP R (“Mediator(+), hnRNP R(+)”). To obtain the relative molar ratios of the transcripts, the measured transcript levels were normalized by the number of C residues within the transcribed region because [α-32P] CTP was used for labeling the transcripts. The molar ratios of the second-round transcripts to the first-round transcripts were calculated using the normalized transcript levels. All the values were averaged from three independent experiments, and error bars represent S.E. The number below each lane indicates the length of the G-free cassette. The p values of the molar ratio (2nd/1st) on each template versus that on the pfMC2AT(350) are as follows. In the presence of only Mediator: p = 0.83 (pfMC2AT300), p = 0.21 (pfMC2AT250), p = 0.16 (pfMC2AT200), p = 0.13 (pfMC2AT150) and p = 0.13 (pfMC2AT100). In the presence of both Mediator and hnRNP R: p = 0.34 (pfMC2AT300), p = 0.042 (pfMC2AT250), p = 0.018 (pfMC2AT200), p = 0.008 (pfMC2AT150) and p = 0.022 (pfMC2AT100).