Mice

Eight-week-old, female, NSG (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ) mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA), with all animal procedures reviewed and approved by the St. Jude Children’s Research Hospital Institutional Animal Care and Use Committee. Mice underwent orthotopic tibial implantation as described below and were followed by weekly bioluminescence. They underwent hindlimb amputation when they reached a humane endpoint that included lameness, large tumor burden interfering with the animal’s ability to reach food or water, significant tumor ulceration, guarding behavior, or upon becoming moribund. After hindlimb amputation, mice were followed with weekly bioluminescence imaging until they reached a total body bioluminescent flux of 1 x 10¹⁰ photons or if other endpoints were seen such as persistent poor grooming or lethargy; > 20% body weight loss post-amputation, respiratory difficulty or upon becoming moribund.