Extended Data Fig. 1: Biochemical analyses of ROOL RNAs.
a, Denaturing PAGE analyses demonstrate that the RNAs are intact full-length molecules. RNAs were analysed at least once. b, Mass photometry analysis of ROOLEfa and ROOLFirm at 50–100 nM in 240 mM K+ and 20 mM Mg2+ shows that monomers and octamers are the two most abundant species. The RNA size of each peak was calibrated using the Millennium RNA Marker and labelled. c, Size exclusion chromatography (SEC) analyses of ROOLEfa and ROOLFirm coupled with denaturing PAGE (d), mass photometry (e), and negative-stain EM (f) showed three major states (aggregates, octamer, and monomer) in the cryo-EM samples. Peak 1 (void volume) contained aggregated ROOL of different oligomerization states; peaks 2 and 3 correspond to predominantly octamers and monomers, respectively. Experiments in c and e were repeated at least once with similar results; experiments in d and f were performed once; three EM micrographs were analysed in f. g, Dynamic light scattering shows a diameter range of 10–30 nm for ROOL RNAs, consistent with its oligomerization states. A representative result from two independent technical replicates is shown. For original RNA gel images and negative-stain micrographs, see Supplementary Fig. 1.
