Extended Data Fig. 6: Preparation and activity of complexes with TFIIF and TFIIF mutants.
From: Mechanism of mammalian transcriptional repression by noncoding RNA
a, Schematic detailing the TFIIF variants used in these studies. b, SDS-PAGE analysis of equimolar amounts of each TFIIF variant. c, Transcription assays show that the TFIIF variants employed in this study have differential ability to stimulate Pol II activity, as previously described47,48. Left, Urea PAGE analysis and phosphorimaging; right, quantification. Data are presented as mean values +/- standard deviation, with individual data points shown as black circles. TFIIF color code as in Fig. 2. d, Transcription assays showing the inhibitory effect of a single-stranded RNA. Only the RNA products well separated from a DNA template-independent RNA product were quantified. Left, Urea PAGE analysis and phosphorimaging; right, quantification. Data are presented as mean values +/- standard deviation, with individual data points shown as black circles. e, Urea-PAGE analysis of transcription assay quantified in Fig. 2c. f, Electrophoretic mobility shift assay (EMSA) showing the binding of wild-type TFIIF and the indicated TFIIF mutants to Alu-LA and Alu-RA. g, Quantification of the fraction of bound RNA monitored by the disappearance of the free RNA band. Results represent analysis of three independently prepared reactions. Data are plotted as mean values +/- standard deviation.
