Nod scid il2rg nsg mice

Animal protocols were approved by the Institution for Animal Care and Use Committee of the University of Southern California. 2.5×10⁶ MOLM-13 cells were administered via tail vein injection into 4- to 6-week-old female NOD-scid /Il2rg^−/− (NSG) mice from Jackson Laboratory (Bar Harbor, ME). Engrafted mice were treated with 200 μL of 220 μM A192 (n=4) or α-CD99-A192 (n=4) via tail vein on Day 7, 10, 13 and 16 post engraftments. Mice were euthanized on Day 21 and spleen, bone marrow and blood were collected and stained for huCD45 and analyzed using flow cytometry. For survival analysis, male mice engrafted with MOLM-13 cells were treated with 200 μL of 220 μM of A192 (n=7) or α-CD99-A192 (n=7) on days 7, 10, 13 and 16 post MOLM-13 engraftment. Mice were euthanized when they displayed signs of severe weight loss, lethargy, hair loss, and/or hunched appearance.

Animal studies were performed in compliance with institutional guidelines under an IACUC approved protocol at MUSC or Arizona State University. NOD-scidIL-2Rg−/− (NSG) mice were obtained from Jackson Laboratory (Bar Harbor, ME) and bred in-house. For the orthotopic mouse model, 25,000 cells derived from LS174T cells were injected into the cecal wall of male NSG mice at the age of 8 weeks. Four days later, PGE₂ (Cayman Chemical #14010) or vehicle were given to mice through intraperitoneal injection at 300 μg/mouse/day. PGE₂ was dissolved in 100% ethanol and diluted to desired concentration with PBS immediately before the injection. The mice were sacrificed at 8 weeks for experiment with p53 overexpression and at 6 weeks for experiment with p53 CRISPR depletion after the start of the treatment. Metastatic nodules on livers were visually inspected and counted. Metastatic nodules on lungs were revealed by India ink staining and counted. Primary cecal tumors were isolated and weighted. Haematoxylin and eosin stain was performed on sectioned liver and lung to confirm tumor histology.

Animal protocols were approved by the Institution for Animal Care and
Use Committee of the University of Southern California. For xenograft
experiment, 4- to 6-week-old male NOD-scid/Il2rg^−/− (NSG) mice purchased from Jackson
Laboratory (Bar Harbor, ME, USA). 10×10⁶ TF-1 cells were
administered via tail vein injection into mice. Mice were randomized into
control and treatment groups. Engrafted mice were treated with 200 μL of
100 nM A192 (n=3) or hGMCSF-A192 (n=3) via tail vein on day 1, 3, and 5 post
engraftments. Mice were euthanized on Day 16 and the bone marrow and blood were
collected and stained for huCD45 (HI30, PE-Cyanine7, eBioscience) and analyzed
using flow cytometry.

All animal work was performed in accordance with institutional guidelines and institutional animal care and use committee (IACUC) approval. We used 5- to 6-wk-old male immunocompromised NOD-SCID IL2Rg^-/- (NSG) mice (Jackson Laboratory) for the study.