Fig. 2: Analysis of species-specific binding patterns.

A Schematic of “control,” “orthologous,” and “bound” oligo classes used for species-specific transcript-level binding analysis. B, C Cumulative distribution function of log₂ nsRBNS enrichment of all iCLIP hits: control (light grey; dotted), orthologous (dark grey), and bound (teal) of (B) CDS and (C) UTR oligos. Inset boxplots show in vitro binding patterns for “bound,” “motif mutant,” and “orthologous” oligos. Significance of inset boxplots was determined via two-sided paired Wilcoxon test (n = 1373 sequences in (B) and 987 sequences in (C)). Significance marks are as follows: ****(p ≤ 0.0001). Centre line denotes median (50th percentile) with bounds of box representing 25th to 75th percentiles and the whiskers denoting 5th to 95th percentiles. Outliers are denoted as individual points. Inset heatmaps show significance values for all comparisons via two-sided KS test and corrected for multiple comparisons via the BH procedure for the cumulative distribution curves. Red denotes significant (p ≤ 0.05). Values are as follows: d (p ≤ 0.01), f (p ≤ 0.0001). D Cumulative distribution function of log₂ fold nsRBNS enrichment change of in vivo bound over in vivo not bound oligos separated by ∆UAG content. Inset shows significance values for all comparisons via two-sided KS test and corrected for multiple comparisons via the BH procedure. Red denotes significant (p ≤ 0.05). Values are as follows: a (ns), c (p ≤ 0.05), e (p ≤ 0.001), f (p ≤ 0.0001). E Cumulative distribution function of RiboSeq fold change, log₂ separated via iCLIP detection. nsRBNS enrichment cutoffs defined as “less enrichment” <1 and “better enrichment” >1. Insets show significance values for all comparisons via two-sided KS test and corrected for multiple comparisons via the BH procedure. Grey denotes nearing significance (p ≤ 0.1). Red denotes significant (p ≤ 0.05). Values are as follows: b (p ≤ 0.1), e (p ≤ 0.001), f (p ≤ 0.0001).