Fig. 2

Indisulam, Pladienolide B and THZ531 modulate specific splicing patterns in 22Rv1 cells. A Pie chart showing % of events regulated in indicated splicing pattern upon indicated treatment in 22Rv1 (fold change ≥ 2, p-value ≤ 0.05). B Bar graphs showing the percentages of events for each splicing pattern annotated in FAST-DB (white columns) or regulated in 22Rv1 upon indicated treatment (black columns, modified Fisher’s test). C Pie charts showing percentages of up- (red) or down-regulated (green) events for indicated splicing pattern (ALE, alternative last exon; EC, exon cassette; IR, intron retention) in 22Rv1 upon indicated treatments. D Venn diagram showing the overlap between annotated splicing events regulated by the indicated in treatment in 22Rv1 compared to control cells. E) Pie chart representing distribution of 35 commonly regulated splicing events by the three treatments in 22Rv1 (see panel D) among different splicing patterns. F-I Representative images of RT-PCR analyses for overlapped (F), Indisulam—specific (G), PladB – specific (H) and THZ531- specific AS events in 22Rv1 cells treated or not for 24 h with indicated drugs (Ind, indisulam; PladB, pladienolide Schematic representation for each event is depicted beside the representative agarose gel. Red and green boxes indicate respectively up- and downregulated exons, black arrows indicate primers used for the PCR analysis. Percentage of spicing inclusion (PSI) was evaluated by densitometric analysis, and results are shown below agarose gels (mean ± SD, n = 3, t-test)