title: Mouse orthotopic transplantation MSA2 12 samples single cell RNA-Seq
library_construction_protocol: Tumours were dissociated to single cell with collagenase D and liberase DL. Cells were stained with antibodies against CD45, Pdpn, and EpCAM. DAPI was used to select live cells Equal number of immune cells, fibroblasts epithelial cells and other cells types were collected by flow cytometry. 15 000 cells were used per lane in the 10x Chromium cartridge.  Mice with tumor diameter ~8-10 mm (8-12 weeks after transplantation) were recruited randomly in MSA-2 arm or the vehicle control arm. The drug regime included oral gavage of 80 mg/kg MSA-2 (MedChemExpress, HY-136927) or vehicle control (50% PEG300 (Sigma-Aldrich, 202371) in 0.9% NaCl) on day 0 and day 5. On day 6, mice were euthanized by CO2, and tumors were harvested and processed for single-cell RNA sequence profiling.  Cells were captured with the 10x chromium v3 protocol without RNA extraction Library were constructed with the 10x Chromium v3 dual index protocol following the manufacturer's instructions