B6 cg tg fabp4 cre 1rev j

The development of conditional PIKfyve^fl/fl mice on a C57BL/6J background by the Cre‐loxP approach was described elsewhere (Ikonomov et al. 2011). To generate mice with selective disruption of Pikfyve in adipose tissue, the PIKfyve^fl/fl progeny was crossed with mice expressing Cre recombinase under the adipose tissue‐specific promoters of aP2 (generated by Ronald Evans and purchased from The Jackson Laboratory; B6.Cg‐Tg(Fabp4‐cre)1Rev/J) and of Adiponectin (a kind gift by Dr. Evan Rosen, supplied by Dr. Richard M Mortensen on a C57BL/6J background). Genotyping was performed by PCR using genomic DNA isolated from tails of 20‐day‐old mice and specific primer‐pairs detailed previously (Ikonomov et al. 2011). PIKfyve^{fl/fl,aP2‐Cre+} and PIKfyve^{fl/fl,Aq‐Cre+} mice were used as homozygous mutants; PIKfyve^fl/fl mice of the respective breeding were used as controls and PIKfyve^{fl/wt,aP2‐Cre+} or PIKfyve^{fl/wt,Aq‐Cre+} were used as heterozygous mutants. Most of the experiments were performed by comparing PIKfyve^fl/fl controls versus the respective PIKfyve^{fl/fl,aP2‐Cre+} and PIKfyve^{fl/fl,Aq‐Cre+} mice. Animals were maintained in a temperature‐controlled environment (22 ± 1°C) with a 12‐h light/dark cycle (6 am/6 pm) and a standard rodent diet (4.5% calories from fat; LabDiets #5001) with free access to food and water. Separate groups of male and female mice were used as indicated.

aP2-Cre mice (Stock number 005069, B6.Cg-Tg (Fabp4-cre)1Rev/J) were obtained from Jackson Laboratory. Generation of floxed GHS-R allele-containing mice has been described previously [20 (link)]. We removed the FRT-PGK-neo-FRT cassette by breeding them with FLP mice, and then backcrossed them for 10 generations onto C57BL/6J background. Mice were bred and housed in a pathogen-free facility at Baylor College of Medicine. Animals were housed under controlled temperature (23 ± 1 °C) and 12 h light-dark cycle with free access to food and water. Normal chow diet (2920X, 16% of calories from fat, 60% from carbohydrates, 24% from protein) was purchased from Harlan-Teklad (Madison, WI, USA). All experiments were approved by the Animal Care and Research Committee of the Baylor College of Medicine. Age-matched male and female wild-type (WT), aP2-Cre, Ghsr^f/f, and aP2-Cre/Ghsr^f/f mice were used in the studies, including young (5–6 months) and old (15–17 months) cohorts. All animal experiments have been approved by Institutional Animal Care and Use Committee at Texas A&M University with the approval code as AUP IACUC 2016-0292.