Genetic Knockout Mouse Model Characterization

C57BL/6 (18–22 week old, male:female ratio 1:1, body weights 20–25 g; Charles River Laboratories, Wilmington, MA), Pld1^−/− and Pld2^−/− were housed in isolation cages in pathogen-free conditions on a light-dark cycle with light from 7:00 to 20:00 at 25°C. Pld1^−/− and Pld2^−/− were obtained from Dr. Gilbert Di Paolo (Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, New York)^{28 (link)} and Dr. Yasunori Kanaho (Department of Physiological Chemistry, University of Tsukuba, Tsukuba, Japan).^{29 (link)} The Pld1^−/− and Pld2^−/− mice were backcrossed with C57BL/6 mice for > 7 generations.^{29 (link),30 (link)} Mice were fed a standard diet (Laboratory Rodent Diet 5001; PMI Nutrition International, St. Louis, MO) containing 4.5% total fat with 0.3% ω-3 fatty acids and <0.02% C20:4 and were provided water ad libitum.