Fig. 4
HNRNPH controls RON exon 11 splicing via multiple intronic and exonic binding sites. a HNRNPH iCLIP validates HNRNPH binding to predicted splice-regulatory binding sites (SRBS). Bar diagram shows the number of HNRNPH crosslink events from HEK293T cells on each position along the wt RON minigene. HNRNPH SRBS (brown boxes) were assigned to five SRBS clusters (circled numbers). iCLIP data show HNRNPH binding to four out of the five SRBS clusters. b Extending or disrupting G-runs results in opposite splicing effects. Scatterplot shows inverse correlation of median changes in AE inclusion (average of three biological replicates) of all mutations per SRBS that extend or disrupt the G-run (see Methods) with linear regression line. r, Spearman correlation coefficient and associated P value. c Exonic and intronic HNRNPH binding exerts distinct effects on RON exon 11 splicing. Boxplots summarise the change in frequencies of each isoform in MCF7 cells (mean, n = 3) for all G-run-disrupting mutations within HNRNPH SRBS of different clusters (circled numbers). Box represents quartiles, centre line denotes 50th percentile and whiskers extend to most extreme data points within 1.5× interquartile range. Number of considered mutations for each cluster given below. *P value < 0.05, **P value < 0.01, one-sample Wilcoxon test against population mean of zero. d Bar diagram shows the changes in isoform frequency of wt RON minigenes upon HNRNPH KD in MCF7 cells. Error bars indicate standard error of the mean from three biological replicates
