Inducible Genetic Manipulation of Dhps in Mice

Mice containing the floxed Dhps alleles were crossed to MIP1-CreERT (21 (link)) mice to generate Dhps^Δβ mice on the C57BL/6J background. Animals were maintained under protocols approved by the Indiana University School of Medicine Institutional Animal Care and Use Committee. The following primers were used for genotyping Dhps^Δβ mice: 5’-GTAAACTAGAGTTCTGCGATGGGTGG-3’ (forward) and 5’-TCAATCTGGTCATAAGGGCACAGG-3’ (reverse), and were expected to generate a 319 bp band for the wild-type allele and 396 bp for the floxed allele. Mice were crossed to B6.Cg-Gt(ROSA)26Sor^{tm14(CAG-tdTomato)Hze/J} and pancreas tissue was harvested to visualize Tomato expression. Male mice with the desired genotype were then weaned and utilized for all the experiments in this study. Mice were administered 3 daily intraperitoneal injections of 2.5 mg of tamoxifen dissolved in peanut oil at 8 weeks of age. Mice were then allowed to acclimate for 1 week prior to being placed on either a NCD (16% kcal from fat; Research Diets; D12492) or HFD (60% kcal from fat; Research Diets; D12492). Glucose and insulin tolerance tests were performed as described (18 (link)). Mice harboring kinase-dead-PKC-ζ protein in β-cells (β-KD-PKC-ζ) (23 (link)) were placed on either a NCD or HFD at 8 weeks of age as previously described (23 (link)) and euthanized for pancreas tissue after 1 week.