Single cell RNA sequencing on PyMT mammary gland primary tumor and lung metastases

Using scRNA-seq of the PyMT/GPx2 KD versus the PyMT control tumor model, we identified an overlap in cell clustering involving six luminal-like clusters and one basal/mesenchymal-like cluster (cluster 3). Similarly, scRNA-seq of lung metastases derived from the GPx2 KD tumor unraveled several luminal-like clusters and one mesenchymal-like cluster (M-cluster) which was highly homologous to primary tumor cluster 3. PyMT-GPx2 KD tumor cells were GFP-labeled in vitro and then injected into mammary fat pad of mice for incubating 45 days. We then harvested GPx2 KD primary tumor and lungs and generated single cell suspension by FACS sorting from GPx2 KD, and paired lung metastases. 10X Genomics was used to make cDNA library. We then sequenced the samples with Illumina high throughput sequencing.