Illumina NovaSeq 6000 sequencing; Single-cell RNAseq of CD45 positive cells in MC38 tumors - Primary irradiated tumors and secondary non-irradiated tumors - Concurrent treatment with anti-PD-L1
Single-cell RNAseq of CD45 positive cells in MC38 tumors - Primary irradiated tumors and secondary non-irradiated tumors - Concurrent treatment with anti-PD-L1
MC38 tumors harvested from immunocompetent C57BL/6 mice treated with different conditions Mechanic and enzymatic dissociation, and then CD45 positive sorting to obtain CD45+ single-cell suspensions to be sequenced CD45+ single-cell suspensions were stored at a concentration of 106 cells/mL and kept on ice until use. Single-cell suspensions were loaded onto a chromium single-cell chip (10X Genomics™) according to the manufacturer's instructions for coencapsulation with barcoded gel beads at a target capture rate of ~10,000 individual cells per sample. Captured mRNAs were barcoded during cDNA synthesis using the Chromium Next GEM Single-Cell 3' gel beads-in emulsion (GEM), Library and Gel Bead Kit v3.1 (10X Genomics™) according to the manufacturer's instructions. The resulting libraries were prepared in parallel in one single batch for each condition.