Fig. 5

Stable G4 lead to more paused POLRMT. A NMR spectra of reference (red) and mutant (blue) CO1-G4 RNA sequence. The position of a G to A mutation is in bold. B Representative gel showing intensity of fluorescently labelled RNA product from in vitro transcription and DNA loading control. Quantification of normalized fluorescence signal is plotted (n = 6; P < 0.01; t-test error bars = S.E.M). C Representative gel showing intensity of fluorescently-labelled RNA product from in vitro transcription using 7-deaza-G and DNA loading control. Quantification of normalized fluorescence signal is plotted (n = 6; P > 0.05; t-test error bars = S.E.M). D Confocal image showing RHPS4 (green) and MitoTracker (grey) localization in the cytoplasm following 4 and 24 h of RHPS4 treatment. Cells in the upper panels are not stained with MitoTracker. Scale bar is 10 µm. E Immunofluorescence staining for G4 using 1H6 antibody (green) before and 24 h after treatment with RHPS4. Scale bar is 50 µm. Bulk G4 abundance measured by immunofluorescence, expressed as percentage of intracellular area labeled, before and after RHPS4 treatment (n = 9; P < 0.001, t-test). F PRO-seq data from biological replicate experiments showing POLRMT distribution before and after treatment with RHPS4. Y-axis is RPM. RHPS4 results in a shift in POLRMT localization with more polymerase at the 5’ end of the polycistron and decreased POLRMT at the 3’ end of the polycistron. G-rich light strand transcripts are in red and G-poor heavy strand transcripts are in blue