Functional heterogeneity of cancer-associated fibroblasts in pancreatic cancer

Single-cell RNA-sequencing analyses were performed on unfractionated live cell mixtures or sorted fibroblasts from pancreatic tumors of KPC;YFP mice, so as to investigate the functional heterogeneity of cancer-associated fibroblasts in early-stage and late-stage tumors. Unfractionated live cell mixtures or sorted fibroblasts from pancreatic tumors of KPC;YFP mice were analyzed by 10x single-cell RNA-sequencing. Cancer-associated fibroblasts were extracted using the reverse sorting selection of CD45/CD31/YFP triple-negative cells. As an alternative route, aSMA-expressing cancer-associated fibroblasts were sorted by RFP expression from KPC;YFP;aSMA-RFP mice.