Fig. 5: RBM17 suppresses EIF4A2 poison exon inclusion.

a Ranked protein expression change curve 5 days after transduction with shRNAs against RBM17 in K562 cells. Protein names that also have significant RBM17 binding with their transcripts (eCLIP-seq) and have significant alternative splicing changes after RBM17 knockdown (at day 5 by RNA-seq) are displayed (colored in yellow). b–d RT-PCR validation of EIF4A2 (b), RBM39 (c) and EZH2 (d) variants using RNA extracted from K562, HL60 and primary AML cells with and without RBM17 knockdown. n = 3 independent experiments. e Upper: Diagram of EIF4A2 splicing variants and the primers for RT-PCR detection of cryptic exon inclusion. Bottom: IGV plot of RNA-seq data illustrating the EIF4A2 cryptic exon promoted by RBM17 knockdown and anti-RBM17 eCLIP-seq tracks. f qPCR quantification of UPF1 knockdown in K562 cells. n = 3, mean ± SD, two-tailed Student’s t test. g EIF4A2 intron 10-included isoform in K562 cells with and without UPF1 knockdown and actinomycin D treatment. n = 4, mean ± SD, two-tailed Student’s t test. h EIF4A2 intron 10-excluded isoform in K562 cells with and without UPF1 knockdown and actinomycin D treatment. n = 4, mean ± SD, two-tailed Student’s t test. i Western blot detection of EIF4A2 after RBM17 knockdown in K562 and HL60 cell lines. n = 3 independent experiments. ***P < 0.001, ****P < 0.0001. Source data are provided as a Source Data file.