Figure 1. Quantification of splicing events by conventional RT‐PCR. Example of primer location for quantification of (a) cassette exon (blue), (b) mutually exclusive exons (blue or purple), or (c) alternative 3′‐terminal exons (green or blue). The black arrows represent the primer location; the primer that is 5′‐end labeled with [γ‐ ³²P]‐ATP is marked with ○. Below each panel is a representative gel showing the outcome of the splicing reaction.

Figure 2. Quantification of alternative splicing changes by real‐time PCR. (a) Schematic representation of a cassette exon with location of specific primers to assess levels of exon inclusion (light blue), exon skipping (dark blue), or gene expression (green); (b) The formula used for the fold change of exon inclusion/skipping in the analyses; (c and d) Representative amplification plots and melting curve analyses, respectively, for each of the primer pairs. Red represents control samples, light blue siRNA 1 samples, and black siRNA 2 samples; (e) Agarose gel electrophoresis carried out to ensure a lack of nonspecific amplification on the qPCR; (f) Histogram showing the changes of inclusion of a cassette exon upon PTB/nPTB knock‐down, assessed with two different sets of siRNAs. The color‐coding is the same as for panels (c) and (d).