Balb c nude

The CRC cell lines MC38, CT26, and HT‐29 were purchased from the American Type Culture Collection (ATCC) and cultured according to the manufacturer's instructions. These cells were characterized using short tandem repeat markers by Genewiz, Inc. and were confirmed to be mycoplasma free (last tested in 2017). Note that 5‐ to 6‐week‐old BALB/c nude, C57BL/6, BALB/c, NOD/SCID, and NOD‐Prkdc^em26Cd52IL2^rgγem26Cd52/Gpt (NCG) mice were purchased from Gempharmatech and were housed under pathogen‐free conditions, in temperature‐controlled holding rooms (20−22°C), with 30%−34% humidity, and on a 12 h on (6 a.m.) and off light cycle (6 p.m.).

The animal studies were approved by the Institutional Animal Care and Use Committee of the First Affiliated Hospital of Nanjing Medical University, Nanjing, China. Five–six-week-old female nude mice (BALB/c-nude) were purchased from GemPharmatech Co., Ltd. (Nanjing, China). In total, three groups with six mice per group were used for the subcutaneous tumorigenesis models. PANC-1–Vector, PANC-1–SOX2OT, and PANC-1–SOX2OT–FUS cells (1 × 10⁶ cells/100 μl) were subcutaneously injected into the subaxillary fossa of right anterior limb in each mouse, separately. When a tumor was palpable, body weight and tumor volume were measured twice weekly. The tumor volume was calculated using the formula (width² × length)/2. About 9 weeks (61 days) later, the mice were euthanized. After the mice were photographed, the subcutaneous tumors were taken, weighed, and photographed. Three groups with six mice per group were used for the tail vein metastasis model, PANC-1–Vector, PANC-1–SOX2OT and PANC-1–SOX2OT–FUS cells (5 × 10⁶ cells/100 μl) were separately injected into the tail vein of each mouse. Four weeks later, the mice were euthanized and lungs and livers were removed. Each tissue sample was cut in two, one was snap-frozen in liquid nitrogen until use and the other was and fixed in 4% paraformaldehyde (for immunohistochemistry).