Freshly harvested tumors from C57BL/6 mice were pooled separately into 4 treatment groups to maximize the isolation of tumor infiltrating  cells. Tumors were gently dissociated using the gentleMACS Octo Dissociatorwith Heaters (Miltenyi Biotech). Single-cell suspensions of tumor cells were purified by incubation with CD45 Microbeads (Miltenyi Biotec, cat. no. 130-110-618) as per the manufacturer's instructions to enrich the tumor-infiltrating  cells. Quality and quantity of cells were assessed under the microscope by trypan blue staining, and dead cells were removed using the Dead Cell Removal Kit (STEMCELL, cat. no. 17899) as per the manufacturer's instructions. Nuclei acid extraction was performed following the manufacturer's instructions (10x Genomics, as part of the Chromium Single Cell 3' v3.1). To capture approximately 7,000 targeted cells, 7,000-8,000 live cells were loaded onto the Chromium Controller (10x Genomics), and scRNA-seq libraries were generated using ChromiumNext GEM Single Cell 3' Reagent kit v3.1 (10x Genomics) according to the manufacturer's protocol.