Extended Data Figure 6: Ribo-seq of unc-54(cc3389) shows an unexceptional progression of ribosomes in the readthrough region.

a, Raw ribo-seq reads for unc-54(+) (blue) and unc-54(cc3389) (green) animals, plotted as read pile-ups. Mismatched bases are indicated with black bars. Location of the normal stop codon and the first in-frame stop codon are indicated with ‘TAA’ and dotted lines. The extension in unc-54(cc3389) is 30 amino acids. b, The number of ribo-seq reads in the last 30 codons, compared to the previous 30 codons, for all mRNAs. Linear regression was performed on all points (solid line), and twofold difference shown (dashed lines). c, The distribution of ribo-seq reads in the last 30 codons (90 nt) of unc-54(cc3389) is shown in green, and the 95% confidence interval (CI) for all open reading frames in dashed lines. d, The fraction of in-frame ribo-seq reads in the last 30 codons is plotted as a function of read counts in the last 30 codons, and unc-54(cc3389) highlighted. e, The distribution of read lengths in the last 30 codons of unc-54(cc3389), and all open reading frames (95% confidence interval, dashed lines). For b–d, reads were restricted to 28, 29, 30 nt lengths. For b–e, a 12 nt offset was performed for the ribosomal P-site, and read counts were derived solely from the unc-54(cc3389) ribo-seq library. For c and e, a minimum 15 read counts was imposed to obtain the 95% confidence interval from ‘all genes’.