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. 2005 Jul 19;33(13):4007–4015. doi: 10.1093/nar/gki710

Figure 1.

Figure 1

Effect on pre-mRNA splicing of modified U1 snRNAs binding along the entire ATM Δ. (A) Schematic representation of ATM hybrid minigenes used in transient transfection assay. Exons are indicated as black boxes, introns as line and dotted lines represent possible splicing products. Primers used in RT–PCR analysis are shown as black arrows. The position where the ATM intron 20 sequences were cloned is indicated. (B) The upper panel shows the relevant portion of the nucleotide sequences of the ATM Δ intron 20. The cryptic exon is shown in uppercase, intronic sequences are in lowercase and the cryptic acceptor (ag) and donor (gc) sites are underlined. The empty underlined space in the sequence corresponding to the GTAA deletion. The lines above the sequence show the position of modified U1 snRNAs binding site. Lower panel shows the result of the splicing assay. pATM Δ minigene was transfected in Hep3B cells alone or with the indicated U1 snRNAs and the resulting RT–PCR products analyzed on 1.5% agarose gel. The lower and higher MW bands correspond to normal intron processing and inclusion of the 65 bp cryptic exon, respectively.