Multigenerational Obesity Paper
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ECM characterizations were performed by staining whole tissues with H&E and Trichrome staining and SHG imaging decellularized and delilpidized ECM. Analysis was completed using imagej software to measure adipocyte size, collagen linearity, and collagen thickness. Migration assays were performed by seeding cells onto decellularized and delipidized ECM, then allowing 24 hours for attachment. Seeded ECM was imaged every 15 minutes for 10 hours. These images were then used to track cell motility using imagej tracking by hand. For invasion assays, cells were seeded onto ECM then primed in the environment for one week. Seeded ECM was then placed into transwell invasion wells and allowed to invade using a 15% FBS gradient. After five days, cells that had invaded through were counted. MMF-ECM were prepared by seeding mouse mammary fibroblasts onto glass slides coated with fibronectin in 12 well plates. They were treated with different concentrations of lepton. Ascorbic acid was added every other day to the media to promote ECM growth. The ECM were collected after three weeks and decellularized before performing SEM and migration assays.