GSM5501832: MDA-MB-231 cells no mitotransfer (mito-mEmerald-), macrophages with mito transfer (mitoRFP+), rep1; Homo sapiens; RNA-Seq
Macrophage/cancer cell cocultures were harvested for flow cytometry using trypsin and subsequently stained with BV711-CD11B. Macrophages were FACS-isolated based on CD11B and mEmerald expression, and further isolated for the presence (RFP-positive) or absence (RFP-negative) of cancer cell mitochondria. Cancer cells were FACS-isolated based on RFP expression, and further isolated for the presence (mEmerald-positive) or absence (mEmerald-negative) of macrophage mitochondria. Following FACS isolation, cells were counted and checked for viability on a hemocytometer using trypan blue. Due to the low efficiency of mitochondrial transfer, each resulting population consisted of low cell numbers. Therefore, we pooled cancer cells that received macrophage mitochondria with macrophages that did not receive cancer cell mitochondria; and pooled cancer cells that did not receive macrophage mitochondria with macrophages that did receive cancer cell mitochondria. Libraries were prepared using the 10X genomics Single Cell 3' Gene expression Library V3 following the manufacturer's protocol.