Fig. 5: His methylation regulates the binding and splicing activity of U2AF1.

a Distribution of changed splicing events when CARNMT1 is deleted, blue for down-regulated and red for up-regulated. b Distribution of changed splicing events when U2AF1 is knocked down, blue for down-regulated and red for up-regulated. c Recovery experiments by supplementing either wild-type or mutated CARNMT1 into CARNMT1 KO cells. GST: Flag-GST, WT: Flag-CARNMT1, M1: Flag-CARNMT1 (D316A), M2: Flag-CARNMT1 (F313A/D316A). 2 times each experiment was repeated independently with similar results. d Two representative altered splicing events were subjected to recovery experiments and quantification of exon inclusion ratio. The data represent mean ± SD. P-value, one-tailed paired t-test. (n = 3 biological replicates). Source data are provided as a Source Data file. e Scheme for illustrating the binding between U2AF complex and pre-mRNA. f Volcano plot showing the alteration in inclusion level for the U2AF1 regulated splicing events (SE events grouped by the nucleotide ahead AG sites) when CARNMT1 is deleted. Source data are provided as a Source Data file. g Altered splicing events of mini-gene expressed in either CARNMT1 WT or KO cells and quantification of exon inclusion ratio. The data represent mean ± SD. P-value, one-tailed paired t-test. (n = 3 biological replicates). Source data are provided as a Source Data file. h eCLIP result for the RNA binding landscape of U2AF1 in CARNMT1 KO and WT cells. i)The binding free energy between U2AF1 and RNA. j MST results (EC50) for the measurement of in-vitro binding between U2AF1 and RNAs. k Structure for illustrating the binding between ZF of U2AF1 and 3’ss of pre-mRNA.