Fig. 5.

Cet1-Ceg1 facilitates promoter escape and assembly of a follow-on PIC for reinitiation. Transcription was initiated with the (A and B) G-less 49 or (C and D) G-less 27 template DNA in the presence (A and C) or absence (B and D) of Cet1-Ceg1 and Abd1. All reactions received 2 mM AMP-PNP prior to 10 to 40% glycerol gradient sedimentation. The gradients were fractionated, and ∼180 µL per fraction was isolated prior to analyzing RNA by denaturing PAGE. The levels of RNA in each fraction were quantified by ImageJ and normalized to fraction 1, and the relative RNA levels were plotted.