Diet-induced obesity mouse model

Sixteen female C57BL/6J mice (n=8/group) were purchased from The Jackson Laboratory (JAX) and ovariectomized at 4 weeks of age. At the age of 5 weeks, mice were randomly assigned to be fed a 10% low fat diet (LFD, 12450Bi) or a 60% high fat diet (HFD, D12492i) for 17 weeks. All mice were sacrificed at 22 weeks of age. Mouse mammary fat pads were obtained and fixed in 4% neutral buffered formaldehyde. Fixed tissue sections were used to assess the levels of p53, HIF1α, PKM2 and aromatase in adipose stromal cells using immunofluorescence. Sample size was selected based on investigators’ previous experience in conducting animal studies. Mouse experiments were conducted in accordance with protocols approved by the Institutional Animal Care and MMaterial Use Committee at Weill Cornell Medicine (New York, NY). Investigators were not blinded.