EO771 subcutaneous tumor_batch 2
Cells from murine tumors were processed using GentleMACS. After red blood cell lysis, tumor cells were stained with Aqua Live/Dead dye (1:150) and incubated at 4 C for 30 minutes. After washing with PBS, cells were resuspended in 1 mL PBS, and 3-5  10 viable cells were sorted for each sample. Cells were then fixed using the 10x Genomics kit according to the manufacturers instructions. Gene expression of fixed samples was evaluated using Chromium Fixed RNA Profiling (Gene Expression FLEX). Samples were hybridized with barcoded probes, and sequencing libraries were generated according to the User Guide (CG000527). Sample concentrations were measured using the automated cell counter Luna-FX7. Samples were then incubated at 42 C overnight (22.5 hours). After hybridization, samples were pooled, and cells were washed and filtered according to the User Guide. The cell concentration of the multiplexed pool was determined before loading onto the Chromium X chip, with a target recovery of 1.28  10 cells. Libraries were sequenced on the Illumina NovaSeq X Plus system according to the manufacturers instructions. FASTQ files were processed using Cell Ranger v8.0.1 (10x Genomics) with the mouse reference genome mm10 (2020-A).