library_construction_protocol: Freshly harvested orthotopic allograft tumor tissues from mice (KPC-shV, n=1; KPC-shZEB1, n=1) were sectioned into 2–4 mm³ segments and temporarily stored in tissue preservation solution (MACS, 130-100-008) at 4°C to ensure simultaneous tissue dissociation and sequencing of all samples. The tissues were carefully washed twice with pre-cooled medium (DMEM supplemented with 0.04% BSA) and thoroughly minced into small pieces. Tissue fragments were then digested at 37°C and 22 rpm for approximately 30 min in an enzyme solution containing DMEM, 0.04% BSA, 0.2% collagenase II (Gibco), and 100 μg/L DNase I (Applichem). Additionally, 0.25% trypsin (Gibco) was added for further digestion for 5 min. The resulting cell suspension was filtered through a 40 μm cell strainer, followed by centrifugation at 300 × g for 7 min at 4°C. MACS Red Cell Lysis Solution (130-094-183) was applied to lyse erythrocytes. Cell pellet after washing with medium was resuspended in 200μL PBS with 1% BSA. The Luna Fluorometer (Logos Biosystems) was used to confirmed the number of viable cells, and the cell suspension concentration was then adjusted to 700-1200 cells/μL. According to the user manual (CG00315) for the 10x Genomics Chromium Next GEM Single Cell 3ʹ Kit v3.1 (1000268), the single cell suspension was immediately loaded onto a chip to generate GEMs (Gel Bead-in-Emulsion) droplets using the 10x Chromium Controller. Reverse transcription, cDNA amplification, and DNA library construction were performed sequentially according to the protocol. The concentration and fragment size of the libraries were measured using the Invitrogen Qubit 4.0 and Agilent 4150 TapeStation. High-throughput sequencing was conducted using high-throughput paired-end 150 bp (PE-150) mode.