Single-cell RNA-seq analysis was performed on CD45-positive cells isolated from mouse mammary tumors.

Tumors derived from BP cells of mouse mammary cancer were collected after treatment with vehicle control, olaparib alone, or the combination of olaparib and an IRG1 inhibitor. CD45-positive cells were isolated from these tumors for single-cell RNA-seq analysis to investigate the impact of the combination therapy on the tumor immune microenvironment. Overall design: Tumor tissues collected from mice treated with vehicle, olaparib alone, or the combination of olaparib and IRG1-IN-1 were enzymatically dissociated into single-cell suspensions using a collagenase/hyaluronidase-based digestion buffer. CD45 positive immune cells were then enriched using a CD45 positive selection kit (18945, StemCell Technologies). For each treatment group, single-cell suspensions from four individual tumors were pooled by centrifugation and resuspended in CryoStor CS10 (100-1061, StemCell Technologies) for preservation. Single-cell RNA sequencing was performed by Novogene using the 10x Genomics platform.