Single cell RNA sequencing of CD45+ cells sorted from tumor tissues of MC38 allograft-bearing mice treated with vehicle or RES

MC38 tumors, treated with either vehicle or RES, were harvested and prepared into single-cell suspensions. CD45+ immune cells were isolated through positive selection using biotin-conjugated anti-mouse CD45 antibody (BioLegend) and MojoSort Streptavidin Nanobeads.The BD Rhapsody Express Single cell Analysis System (BD Biosciences) was employed for single cell capture and cDNA reverse transcription. The size distribution of the cDNA libraries (250-1000 bp) was assessed using the DNA 1000 assay on the Agilent 2100 system (Agilent Technologies). Sequencing was conducted on a NovaSeq 6000 (Illumina) platform.