Fig. 1

Generation of conditional Stac3 knockout mice. a Schematic representation of the process of generating conditional Stac3 knockout mice. The original mutant Stac3 allele contained a “trapping cassette” between Stac3 exons 1 and 2 and a loxP site between exons 5 and 6. Crossing heterozygous Stac3 mutant mice with mice expressing the Flp recombinase caused the trapping cassette to be deleted at the two FRT sites and hence converted the mutant Stac3 allele to a floxed allele, i.e., a pre-conditional wild-type allele. Crossing mice bearing the floxed Stac3 allele with mice expressing the Cre recombinase caused exons 2 to 5 flanked by the loxP sites to be deleted and hence inactivated the Stac3 gene. In the Cre recombinase transgenic mice used in this study, the Cre gene is located downstream of a tamoxifen-inducible chicken beta actin promotor. Arrows labeled with F1, R1, F2, and R2 indicate the locations of PCR primers for genotyping. b Representative gel images of genotyping. Genotypes of Stac3 ^fl/fl, Stac3 ^+/fl Tg ^Cre, and Stac3 ^fl/fl Tg ^Cre mice before and 4 weeks after tamoxifen injection were identified by PCR of genomic DNA followed by gel electrophoresis. Names of PCR primers or target genes are indicated on the right of gel images and sizes of expected PCR products on the left of gel images. WT wild type