Chromium Next GEM Single Cell 3’ RNA-sequencing of tumor cell line co-cultured with PBMC and treated with trifluridine or DMSO control

Chromium Next GEM Single Cell 3’ RNA-sequencing. HCT116 were seeded in 6-well Nunc plates (50,000 cells/3mL/well) and precultured for 24 h before PBMCs were added at a 1:8 ratio. Co-cultures were treated with DMSO vehicle (0.1%) or FTD (3mM) for 12 h or 72 h. MACS Dead Cell Removal Kit (Miltenyi Biotec, Gladbach, DEU) was performed according to the manufacturer’s instructions on cells treated for 72 h to increase the viability of the samples before RNA-sequencing. The viability of the samples treated for 12 h was not subjected to Dead Cell Removal as the viability was already sufficient. All samples were washed in PBS with 0.04% BSA (2x1mL). Chromium Next GEM Single Cell 3’ library preparation and RNA-sequencing were performed by the SNP&SEQ Technology Platform (National Genomics Infrastructure (NGI), Science for Life Laboratory, Uppsala University, Sweden).