GSM9518424: Dnmt1iECKO EC; Mus musculus; RNA-Seq
YUMMER1.7 tumors from control and Dnmt1iECKO mice were harvested and enzymatically dissociated into single-cell suspensions using collagenase II, dispase, and DNase I, followed by mechanical dissociation. Red blood cells were lysed and cells were stained with viability dye and antibodies against CD31 and CD45. Live CD31⁺CD45⁻ tumor endothelial cells were sorted by flow cytometry, washed, and resuspended in PBS supplemented with 0.04% BSA prior to single-cell library preparation. Single-cell libraries were prepared using the 10x Genomics Chromium Single Cell 3′ Gene Expression GEM-X v4 kit according to the manufacturer's instructions. Briefly, sorted cells were loaded onto the Chromium Controller for partitioning into GEMs, followed by reverse transcription, cDNA amplification, and library construction. Libraries were sequenced on an Illumina NextSeq 2000 using the P2 XLEAP-SBS 100-cycle kit in paired-end mode, targeting approximately 20,000–25,000 reads per cell.