Nod cg prkdc il2rg szj

Tissues were obtained from consented patients with Stage II–IV CRC who had undergone surgery at UK Medical Center (IRB #16-0439-P2H). 6–8-week-old NSG mice (NOD.Cg-Prkdc Il2rg /SzJ) from The Jackson Laboratory (Bar Harbor, ME) were used for PDX models. All procedures were performed using protocols approved by the UK Animal Care and Use Committee. Briefly, CRC tissues (2–5 mm) obtained from CRC patients of both sexes were implanted subcutaneously into their flanks in a small pocket surgically created under the skin. Established tumors were designated as generation 0 (G0). Tumor tissues from G0 were minced and mixed with Matrigel to ensure homogeneous distribution of tissues among mice and allow implantation of an equal volume of tumor tissues into the flank. Tumor tissues were resected when they reached an appropriate size and digested as previously described (1 (link)). For evaluation of CD36 expression in PDX models, we utilized tissue samples from Pt 2402 PDX model established from a patient diagnosed with metastatic adenocarcinoma (lung) consistent with colon primary tumor (1 (link)). Pt 2402 PDX tumors were grown to ~200 mm³ then tissues were collected and lysed for analysis via western blot.

The five-week-old NSG (NOD.Cg-Prkdc Il2rg/SzJ) mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA) to generate a GBM intracranial xenograft model. A total of 5 × 10⁵ of U87-FLuc cells were stereotactically injected into mice using the Stereotaxic Instrument (Fisher Scientific) following our reported protocol [39 (link)].

All animal experiments were performed in female, 6- to 8-week-old immunodeficient NSG mice (NOD.Cg-Prkdc Il2rg/SzJ, The Jackson Laboratory, ME), in accordance with the principles and procedures outlined in the NIH Guide for the Care and Use of Animals and approved by the Animal Care and Use Committee of the National Institute of Health (Protocol NOB-008). Cells were subcutaneously injected with (5 × 10⁶ cells in 100 μl HBSS for each mouse) in the right flank. Xenograft size was measured with a sliding caliper twice a week, according to the formula: TV = (width)² × (length)/2. When tumors reached a minimum of 100 mm³, animals were randomized into two groups: vehicle controls and CB839 treatment. Following the treatment with or without CB839 (160 mg/kg, oral gavage, twice a day), the mice were euthanized, and the tumor xenografts were harvested 4 h after the last treatment. The tumor tissues were then divided for two parts. One was reserved in − 80 °C immediately for metabolomics, and the other was fixed with 4% paraformaldehyde for histological and immunochemistry analysis.