Fig. 3
Core and σ70 mutants affect ITC6 pause exit probability. a Experimental conditions studied here and schematic of the different RP complex variants, all with ApA starting substrate. The same experiments have been performed with ATP starting substrate and are presented in Supplementary Fig. 2e, f, g. b Bulk reaction performed with a consensus lac promoter (Supplementary Fig. 1a) and different holoenzyme mutants, in ITC11 NTP conditions (500 µM ApA and 80 µM ATP/UTP/GTP or 500 µM ATP and 80 µM UTP/GTP) and 1 min incubation at 37 °C (the experimental procedure is described in the Supplementary Information). The band assignment is indicated on the left hand side of the gel. c The fraction of transcriptionally active RPs, which displayed NTP-dependent EFRET changes, of all surface-immobilized RPs (see also, Methods: FRET pair localization and detection), in the experimental conditions described in a. d ITC6 pause exit rate for the experimental conditions described in a. e Probability to reach the FS FRET level on the first attempt for the experimental conditions described in a. Error bars are either 1 SD from 1000 bootstraps procedure d or 95% confidence interval c, e
