Fig. 3. Generation of mutant mice lacking MFCS1. (A) Design of the targeting vector. A neomycin (Neo) cassette used for positive selection was flanked by a 5652 bp long arm (AB114903 98833-104485) on the 5' end, a 2357 bp short arm (AB114903 95309-97666), and a diphteria toxin (DT) cassette used for negative selection on the 3' end. Homologous recombination results in deletion of 1167 bp of MFCS1. (B) PCR primers p1 and p2 were used for screening recombined ES clones. (C) Genotyping of mice was carried out using the p3 and p4 primers for the wild-type allele, and (D) the p5 and p6 primers for the MFCS1 KO allele. Ba, BamH1; N, Not1; H, Hpa1.