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. 2006 Jul 19;34(12):3421–3433. doi: 10.1093/nar/gkl390

Figure 4.

Figure 4

Quantitative analyses of the effect of stop codon removal on latent splicing. RNA isolated from cells transfected with the indicated CAD constructs were analyzed by real-time PCR using the specific primer pairs. (a) Gel electrophoresis of the real time PCR products with the latent-specific primer pair. No band is detected in transfections with CAD1 (lane 1) or CAD2 (lane 3), whereas the mutated constructs lacking the intronic stop codons gave rise to a single band of the expected size (96 bp; lanes 2 and 4, respectively). NTC, no-template-control. (b) Quantitation of the real-time PCR results. Fluorescence reading from the ‘authentic/b’ primer pair was defined as 1.00 and all other readings were scaled accordingly. Note that latent splicing in the wild-type constructs, CAD1 and CAD2, is fully suppressed. Error bars represent SE of three independent experiments.