2.1. Animals

To selectively disrupt the BBSome in endothelial cells, we crossed Bbs1^fl/fl female mice with Tie2^Cre male mice, in which Cre recombinase is driven by a pan-endothelial-specific Tie2 promoter [18] (Strain: B6.Cg-Tg (Tek-cre)1Ywa/J; The Jackson Laboratory, stock no. 008863). To visualize Cre recombinase, we first crossed the Tie2^Cre mice with the ROSA (tdTomato) reporter transgenic mouse line, in which a stop codon flanked by loxP sites precedes the start position of a tdTomato locus (Strain: ROSA [Stop^fl/fl-tdTomato]; The Jackson Laboratory, stock no. 007914). Cre recombination removes the stop site, leading to the expression of the fluorescent tdTomato protein. All mice were maintained on the C57BL/6 J background for this study. The age at which mice were studied is indicated in the description of the results of each experiment.

Genotyping of the mice was performed using a polymerase chain reaction (PCR) assay, as described previously [4]. Animals were housed at the University of Iowa vivarium in a 23 °C temperature-controlled environment with a 12-h:12-h light–dark cycle (lights on: 6 AM–6 PM), with ad libitum access to tap water and either standard chow or a high-fat/high-sucrose (HFHS) diet (Research Diets Inc, {"type":"entrez-nucleotide","attrs":{"text":"D12331","term_id":"2148494","term_text":"D12331"}}D12331). Animal procedures were approved by the University of Iowa Institutional Animal Care and Use Committee.