Single-cell RNA-seq of MC38 tumor model
Single-cell RNA-seq of MC38 tumor model
We sorted live, CD45+ cells from single-cell suspensions of tumors from 3 mice using a FACSAria (BD Biosciences). Additionally, T cells from 4 mice were isolated (live, CD45+, TCRb+). Gene expression and TCR libraries were generated using the Chromium Single Cell 5′ Library and V(D)J Reagent Kit (10X Genomics) according to manufacturer's recommendations. 20,000 cells per sample were loaded into each channel of the Chromium Chip, and recommendations were followed assuming targeted cell recovery of 2,000–10,000 cells. 10X Genomics single-cell gene expression libraries were quantified with the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and profiled using the Bioanalyzer High Sensitivity DNA Kit (Agilent Technologies).