FIG 7.

Target genes of WT and mutant SRSF2 by RASL-seq analysis. (A) Unsupervised hierarchical clustering showing that splicing events of WT, P95H, and Δ8aa (Δ8) cells in the absence of doxycycline (Dox −) are very similar and are separated from those of cells with activated shRNA/SRSF2 expression (Dox +) (left panel). Δ8aa and P95H data are grouped side by side, with Δ8aa showing a stronger signal. Results of unsupervised hierarchical clustering of splicing events consisting of shRNA-only cells cultured in the presence or absence of Dox are also shown (right panel). Green, less short isoform, more long isoform; red, more short isoform, less long isoform. (B) Venn diagram showing overlaps and differences of splicing events upon Dox-activated expression of WT, P95H, and Δ8aa SRSF2 compared to shRNA-only expression. (C) Venn diagram showing significant overlapped events (n = 487) from the P95H and Δ8aa groups relative to the WT SRSF2 group. Among these shared events, over 96% were oriented in the same direction. For the significant events that changed in the same direction, Δ8aa-induced changes in 450 of 470 events were over 1.5-fold greater than those corresponding to P95H-induced events. (D) Validation of 10 selected RASL-seq events. Upper panel, heat map demonstration of the ratios of 10 RASL-seq events (short isoform/long isoform). Green, less short isoform, more long isoform; red, more short isoform, less long isoform. Bottom panel, RT-PCR validation of 10 RASL-seq events.