Expression of BRCA2 Δexons 4-7 transcript in human cell lines carrying IVS7 + 2T > G or another truncation mutation at exon 7. (A) RT-PCR analysis of different human cell lines: GM05920B (+/+ LCL), AVO35 (+/IVS7 + 2T > G LCL), AC389 (c.3827delGT/IVS7 + 2T > G fibroblast), SB1685CB (c.3827delGT/IVS7 + 2T > G leukemia), and GM14805 (+/W194X LCL). (B) RHCglo minigene reporter construct is shown in the left panel. BRCA2 exon7 variants replaced the RHCglo exon. Wild-type (WT) and W194X (c.809G > A) mutated were PCR-amplified using overlapping oligonucleotides containing WT or mutated sequences and BamHI and XhoI flanking restriction sites. Primers used for RT-PCR are marked by arrows and indicated as pF and pR. RT-PCR analysis of BRCA2 exon 7 alternative splicing in RHCglo minigene is shown at right. (C) Expression of Δexon 7 and Δexons 4-7 transcripts in ES cells carrying the W194X BRCA2 transgene. (D) WB showing the expression of BRCA2^{Δexons 4-7} variant in W194X ES cells. Actin was used as a loading control. (E) Methylene blue staining of HAT^r colonies obtained after Cre expression into the ES cells expressing either WT or W194X BRCA2.