An ESRP-regulated splicing program is abrogated during the Epithelial Mesenchymal Transition

Figure S1. Additional validations of microarray predicted ESRP regulated splicing events. Semiquantitative RT-PCR was used to assay a set of more complex candidate alternative splicing events. Identities of PCR products are labeled to the right of the gels. The percent inclusion of the relevant exon(s), shaded in red, is given below each lane. For the mutually exclusive splicing events in the FYN and MAPK14 gene transcripts, PCR products were analyzed by restriction digest in order to distinguish the different splice variants of similar size (U—uncut; B—BclI; M—MwoI). In these cases, percent Exon 7b or Exon 9a inclusion was calculated as [MwoI resistant band / (MwoI resistant band + BclI resistant band)]. For the mutually exclusive splicing event in the RAB6A gene transcript, PCR products were digested with restriction enzymes PstI (P) or TspRI (T). Percent Exon 4a inclusion was calculated as [TspRI resistant band / (TspRI resistant band + PstI resistant band)]. Primer sequences are listed in the Supplemental Experimental Procedures.