Fig. 2 E2F1 affects alternative splicing of E2F target genes.

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Fig. 2 E2F1 affects alternative splicing of E2F target genes. E2F1 affects alternative splicing of E2F target genes. (A) A heatmap displaying absolute values of ΔΨ (percent spliced in) for each cell line, corresponding to statistically significant alternative splicing event changes to E2F1 target genes (as determined by the presence of ChIP-seq peaks in their promoter and gene regions, retrieved from ENCODE data) with respect to the pTRE empty vector cell line, derived by analyzing the RNA-seq data with rMATS algorithm. Yellow color represents the lowest difference, and blue color represents the highest. Ivory blocks correspond to nonsignificant changes in splicing patterns (FDR > 0.01). See also table S2 and fig. S3. (B) Pie chart showing the percentage of genes identified in the rMATS splicing analysis, which are E2F1 target genes (as determined by the presence of ChIP-seq peaks in their promoter and gene regions, retrieved from ENCODE data) (i). The Venn diagram demonstrates the overlap of E2F1 target genes affected by alternative splicing events (FDR < 0.01) in each cell line (ii). These data were generated from three independent biological samples. (C) Bar chart displaying the statistically significant alternative splicing events to E2F target genes for each cell line, as compared to the pTRE vector control. The percentage of these alternative splicing changes corresponding to different types of splicing event is displayed in different colors. SE, skipped/cassette exon; RI, retained intron; MXE, mutually exclusive exons; A5SS, alternative 5′ splice site; A3SS, alternative 3′ splice site. See also fig. S2B. (D) Venn diagrams showing overlap between E2F1 target genes identified in the differential expression analysis as being up- or down-regulated (regulated greater than twofold; Fig. 1B) and those identified as being differentially or alternatively spliced [(A) and table S2]. These data were generated from three independent biological samples. (E) Bar chart representing the average fold change in expression of differentially expressed E2F1 target genes (regulated greater than twofold), compared with the expression of those E2F1 target genes where alternative splicing occurred. Only 389 genes from the alternative splicing analysis met the significance threshold for differential expression (P < 0.01). The remaining 632 spliced genes had expression levels that were not significant from the pTRE empty vector cell line (P > 0.01) and were therefore assigned an arbitrary value of 1 for this analysis. Alice Poppy Roworth et al. Sci Adv 2019;5:eaaw4640 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).