FIG. 1.

Generation of COX10 knockout mouse cells. (A) loxP sites were introduced into COX10 by homologous recombination of a plasmid containing a floxed exon 6 (box marked “6”) in embryonic stem cells. The diagram shows the position of the loxP sites in the gene (black triangles). A primary culture from skin fibroblasts was established from a homozygous mouse with the floxed COX10 and transfected with the pCre-Hygro plasmid to generate KO cells. To generate KO cells, several hygromycin-resistant clones were obtained and characterized. (B) Genetic characterization of the COX10 locus. The parental fibroblast line (C) and five hygromycin-resistant clones were analyzed by Southern blotting to detect the deletion of COX10 exon 6. The deleted allele was detected in the K clones (K11, K18, and K19); the wild-type floxed allele was detected in two C clones (C12 and C16). Clone C12 showed the presence of both the floxed and the deletion allele, indicating that it was heterozygous for the deletion. DNA was digested with BamHI, and the location of the probe is shown as a bar (P) in panel A.