Lfd d 12450b

Eight weeks old male Asc^−/− mice and their wild type littermates were used in the present study [11 (link), 41 (link)]. The mice were fed either a low fat diet (LFD: D 12450B, 10 kcal % fat, Research Diets, New Brunswick, NJ) or a high fat diet (HFD: D 12492, 60 kcal % fat, Research Diets, New Brunswick, NJ) for 12 weeks [42 (link)–44 (link)]. In another series, eight weeks old male C57BL/6J or DBA/2J wild type mice (Jackson Laboratories, Bar Harbor, ME), Asc shRNA or a scrambled shRNA (Origene, Rockville, MD, USA) plasmid with a luciferase expression vector was co-transfected into the kidneys via intrarenal artery injection using the ultrasound microbubble system as we described previously [45 (link)]. After the delivery of plasmids into the kidney, mice were fed either a normal diet or HFD for 12 weeks. All protocols were approved by the Institutional Animal Care and Use Committee of the Virginia Commonwealth University.

All animal procedures were approved by Augusta University’s Institutional Animal Care and Use Committee (IACUC). Mice were maintained under standard conditions with controlled 12-h/12-h light-dark cycle and 21 ± 1°C room temperature. Lep^ob/ob, Lepr^db/db and C57BL/6J mice were obtained from Jackson Laboratory (Bar Harbor, ME). Fam13a^-/- mice were backcrossed to C57BL/6 (Stock#: 027, Charles River) for 7 generations and genotyped as previously reported^{14 (link)}. 4–5 week old Fam13a^+/+ and Fam13a^-/- littermates were fed with high fat diet (HFD, D12492, 60% Kcal from fat, Research Diets, NJ) for up to 12 weeks. Mice were not randomized. Six week old C57BL/6J male mice were simply randomized into two experimental groups for ad libitum access to low fat control diet (LFD, D12450B, 10% Kcal from fat, Research Diets, NJ) and 60% HFD for up to 12 weeks as reported^{16 (link)}. All mice studies were not blinded. All mice were sacrificed after 4 h fasting with isoflurane.