Figure 1

Targeted disruption of Hltf in mice by homologous recombination. (A) Schematic representation of the first 7 exons of the mouse Hltf locus, the gene-targeting vector, and the mutant alleles. The location of the hybridization probes (5' probe and 3' probe) for Southern blot analyses are shown. (B) Southern blot analysis of targeted ES cell clones using 5' probe. The genomic DNA was digested with Bam H1 and hybridized with the 3' probe, giving a wild-type allele of 8.7 kb and a mutant allele of 5.6 kb. (C) Southern blot analysis with the 3'probe, showing the targeted allele with a size of 8.8 kb and the wild-type allele of 12.1 kb fragment upon Eco RI digestion. (D) Northern blot analysis of Hltf transcripts in the E10.5 embryos of the indicated genotypes using cDNA probes that cover 5' and 3' coding sequence of Hltf, respectively. The 28S and 18S ribosomal RNAs are shown as loading controls. (E) Real-time RT-PCR for detecting the 3' coding sequence of Hltf in derived Hltf ^-/- ES cells. The presented relative expression was normalized to the expression of Gapdh. No Hltf transcripts were detected in Hltf ^-/- samples by both Northern blot and real-time RT-PCR assays.