Ivis lumina xr 3

This work obtained the four-week-old male nude mice in Model Animal Research Center of Nanjing University. The experimental was carried out in the specific pathogen-free environment in Laboratory Animal Center of Jiangsu University. Each experiment was carried out in line with specific protocols as well as internal biosafety and bioethics guidelines from Jiangsu University and the Jiangsu Municipal Science and Technology Commission (SYXK(su)2018-0053). Approximately 2×10⁶ MGC-803 cells (negative control, sh-circPGD, and sh-ABL2 cells; n = 4/group) per mouse were subject to resuspension within sterile PBS before injection to nude mouse back. After three-four weeks, each mouse was performed cervical dislocation, followed by collection of tumor tissues for further analysis. For metastasis studies, each nude mouse was given injection of approximately 2 × 10⁶ BGC-823 cells with stable GFP-labeled vector transfection via the tail vein (n = 4/group). Under anesthesia with 2.5% isofluorane, the mice were viewed with the IVIS Lumina XR III in vivo imaging system (PerkinElmer, Waltham, CA, USA) and the metastatic status was analyzed after 4 weeks.

Mice (n = 3) were injected with 1 × 10⁷ IU GFP-NLuc virus via i.p. injection or i.v. injection. After 5 days, the mice were sacrificed and the tissues homogenized in cold PBS at 10% weight/volume in lysing matrix D tubes (MP Biomedicals) with a FastPrep-24 5G homogenizer (MP Biomedicals). The lysate was mixed with an equal volume of Nano-Glo Luciferase Assay Reagent (Promega), and luminescence was quantified on an EnVision 2103 Multi-label plate reader (PerkinElmer). For localization of transduced cells in live mice, infected mice (n = 3–6) were injected i.p. with 100 μL of 1:40 diluted Nano-Glo substrate and, after 3 minutes, imaged on an IVIS Lumina III XR (PerkinElmer).

Female NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac (NOG; Taconic Biosciences; Ejby, Denmark) mice were irradiated with 2.5 Gy using the RS-2000 X-ray source (Rad Source Technologies Inc.; Suwanee, GA, USA) after 3 days of receiving antibiotic-supplemented water. On the same day, the mice were engrafted with 2 × 10⁶ WT, NOX2-KO or luciferase-tagged PLB-985 variant cells by tail vein injection. HDC (1 mg/mouse) was administered intraperitoneally (i.p.) three times weekly starting 2 weeks after transplantation. Animals showing symptoms of disease were euthanized and BM cells and other tissues were harvested for flow cytometry, including analysis of human CD11b along with human and murine CD45 and histopathology. For the bioluminescence studies, tumor progression was monitored weekly on an IVIS Lumina III XR (Perkin Elmer; Waltham, MA, USA) after i.p. injection of luciferin (150 mg/kg) and anesthesia. All animal experiments were approved by the Research Animal Ethics Committee at the University of Gothenburg.