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. 2018 Feb 10;8(3):461–469. doi: 10.1002/2211-5463.12389

Figure 3.

Figure 3

The minigene splicing assay based on the pSPL3 exon‐trapping vector. (A) The pSPL3 vector contains two exons SD and SA, and a functional intron, with transcription beginning following the SV40 promoter and ending at the LPAS (late poly (A) signal). Wild and c.1221A > G mutant CUL3 fragments containing 267 bp of intron 8, 171 bp of exon 9, and 185 bp of intron 9 were separately cloned into the XhoI and NheI cloning sites of the pSPL3 vector. (B) Agarose gel electrophoresis of RT/PCR products. SD6 and SA2 primers were designed for RT/PCR amplification of cDNA sequences generated by transfected 293T and Hela cells. Lane 1: marker; Lane 2: empty vector (263 bp); Lane 3–6 : 434 bp (263 bp + 171 bp) and 263 bp. MCS: multiple cloning sites.