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. 2015 Aug 10;112(34):E4726–E4734. doi: 10.1073/pnas.1514105112

Fig. 5.

Fig. 5.

Exon inclusion/exclusion in minigene reporter assays. Splicing reporter minigenes were cotransfected with plasmid expressing WT (W) or P95H mutSRSF2 (P) into 293T cells. Exon inclusion and exclusion isoforms were examined by RT-PCR. Three independent experiments were performed (n = 3). (A) MELK cassette exon minigene containing the native 10-nt site and its mutant minigenes containing mutated 10-nt sites as indicated. The 5-nt motif sequences were capitalized. Note that a cryptic 3′ splice site was detected in the UUUUU1 minigene (last two lanes). (B) The ATF2 cassette exon minigene containing the native 10-nt site and its mutant minigenes containing mutated 10-nt sites as indicated. The 5-nt motif sequences were capitalized. In the last two lanes, the PCR products of the exon inclusion isoform (indicated by an arrow) appeared to migrate slightly more slowly than the PCR products in other lanes. However, DNA sequencing indicated that the PCR products in the last two lanes had exactly the same 5′ and 3′ splice sites as the PCR products in the first two lanes.