Fig. 6: Changes in the splicing landscape upon Snord67 knockout in 4T1 cells.

A Distribution of types of alternative splicing events (Sig Events) in Snord67KO-1 and Snord67KO-2 cells compared to 4T1 WT cells. CE cassette exon, RI retained intron, A3SS alternative 3′-splice site, A5SS alternative 5′-splice site, MXE mutually exclusive exons. Splicing diagrams created in Adobe Illustrator. B, C Volcano plots of differential CE events with |ΔPSI | ≥ 0.1 and FDR < 0.05 in Snord67KO-1 and Snord67KO-2 cells compared to 4T1 WT cells. Exons with significantly increased inclusion in Snord67 knockout clones relative to 4T1 WT cells are shown in blue, while exons with significantly decreased inclusion in Snord67 knockout clones are shown in red. PSI percent spliced in, FDR false discovery rate. D Venn diagram showing the overlap between genes with differential CE events in Snord67KO-1 cells and Snord67KO-2 cells compared to 4T1 WT cells. E, F Gene ontology analysis of genes with differential CE events in 4T1 WT cells versus Snord67 knockout cells. Enrichment analysis was performed in R using the enrichGO function with a p-value cut-off of < 0.05, and the Benjamini-Hochberg procedure was applied to adjust for multiple testing. Individual p-values are provided in Supplementary Data 4. G Cumulative distribution of intron retention ratio in 4T1 WT cells, Snord67KO-1 cells, and Snord67KO-2 cells. Statistical significance was determined by Kolmogorov-Smirnov test (Snord67KO-1 vs. 4T1 WT, p < 0.0001; Snord67KO-2 vs. 4T1 WT, p < 0.0001).