Fig. 1: Direct visualization of in vitro-generated R-loops using EM.
From: Direct visualization of transcription-replication conflicts reveals post-replicative DNA:RNA hybrids
a–d, Native EM analysis of in vitro-transcribed pFC53. Whenever indicated, samples were linearized and/or digested with 6 U/µg RNase H. At least 70 molecules were quantified per condition and replicate. a, R-loop frequency, shown as mean ± s.d., n = 4 independent biological replicates. Statistical significance was determined by ordinary one-way analysis of variance (ANOVA), followed by Sidak test. b, Representative electron micrographs. Scale bars, 200 nm. c, R-loop size; the median is indicated in red. d, R-loop position (black bar) on linearized pFC53 (gray). e–g, Immuno EM analysis of in vitro-transcribed and S9.6-gold-labeled pFC53. Whenever indicated, samples were linearized and/or digested with 6 U/µg RNase H. At least 70 molecules quantified per condition and replicate. e, S9.6-gold binding frequency as mean ± s.d., n = 3 independent biological replicates. Statistical significance was determined by ordinary one-way ANOVA, followed by Sidak test. f, Representative electron micrographs. Scale bars, 200 nm. g, S9.6-gold binding position (black dot) on the linear pFC53 (gray). Numbers below x-axes in a and e indicate the total number of molecules analyzed in all replicates. In d and g the position of the R-loop forming mAirn gene is indicated in light blue below the graph.
