Figure 6.

Stress-induced alternative splicing networks share common features with RMS tumors. Human Exon 1.0 ST Array was used to examine alternative splicing events induced on cisplatin treatment of Rh30 cells (75 µM for 12 hours). (A) Heat map displaying differential expression of genes between normal and cisplatin-treated Rh30 cells for three biologic replicates. (B) Pie chart displaying the number of genes showing alternative splicing and/or differential expression and those that remain unchanged on cisplatin treatment. The DE grouping contains genes with two-fold changes in expression levels and P < .05 with FDR correction. The AS group contains all genes that show alternative splicing at P value < .05 with FDR correction. (C) Pre-mRNA from three RMS tumors (alveolar A4217 and A1063 and embryonal E2035) that showed the presence of both MDM2-ALT1 and MDM4-ALT2 and normal skeletal muscle tissue (Clontech and Life Technologies) was assayed for the splicing of five targets identified in the exon microarray. All the RMS tumors examined in comparison with normal skeletal muscle tissue showed splicing patterns similar to cisplatin-treated Rh30 cells. (D) Table showing the documented functions of CEP97, PELI1, EED, LSM1, and C13orf23, which were examined for altered splicing in the RMS tumors. The P values indicated in the table represent the significance of the difference in the expression of the cassette exon (Figures 5C and W1) in these genes between normal and stress conditions as determined using the Partek software.