Fig. 3: Indisulam-mediated RBM39 degradation leads to the mis-splicing and depletion of proteins regulating cell cycle and metabolism. | Nature Communications

Fig. 3: Indisulam-mediated RBM39 degradation leads to the mis-splicing and depletion of proteins regulating cell cycle and metabolism.

From: Indisulam targets RNA splicing and metabolism to serve as a therapeutic strategy for high-risk neuroblastoma

Fig. 3

a Volcano plot of proteomic analysis in IMR-32 cells treated with 5 µM indisulam or Vehicle control (VC, 0.1% DMSO) for 16 h. b Overlap of splicing events (exon skipping and intron retention) with up or down-regulated proteins after 16 h of Indisulam. Gene ontology analysis of mis-spliced down-regulated proteins and pathway analysis. c RNA Read counts of TYMS (exon 1–5) in IMR-32 cells treated with VC (top), 1 µM (middle) or 5 µM (bottom) indisulam for 16 h. The red box indicates intron retention between exon 4 and 5. d Diagram of custom primers to detect skipping of exon 2–4 of CDK4. e PCR of CDK4 (exon 1–5) in IMR-32 (top) and KELLY (bottom) cells treated with VC, 1 µM or 10 µM indisulam for 0.5–24 h. Representative blot for n = 2 independent experiments. f Western blot analysis of IMR-32 (top) and KELLY (bottom) following treatment of VC or 5 µM indisulam for 16 h. Membranes probed for RBM39, TYMS, CDK4 and ß-actin (n = 3 independent experiments). g Densitometry analysis of CDK4 in IMR-32 (top) and KELLY (bottom). Data are presented as mean values ± SEM. Statistical significance was determined by a two-tailed one-sample t-test. Source data is provided as a Source Data File.

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