Grad-seq visualizes the RNA/protein complexome of C. difficile. (A) Grad-seq workflow. (B) A260 absorbance profile of gradient fractions. Low-molecular-weight complexes (bulk peak) and ribosomal subunits (30S, 50S) are highlighted. (C) Ethidium bromide stained PAA gel showing gradient distribution of housekeeping RNAs (tmRNA, tRNAs, 5S rRNA, 16S rRNA and 23S rRNA) and RaiA. (D) Coomassie stained SDS gel showing gradient distribution of proteins. Ribosomal subunit proteins (30S and 50S) are highlighted. (E) Heatmaps showing normalized sedimentation profiles of selected housekeeping RNA-protein complexes. Data were obtained by RNA-seq and LC-MS/MS analysis of gradient fractions. Sedimentation profiles of proteins and transcripts are scaled in the range from 0 to 1. Fractions 1–20 and the pellet (P) are shown. *not available in previous genome annotations.