GSM7789682: shRen luminal rep4; Mus musculus; RNA-Seq
Mammary gland tissue was dissociated using gentle collagenase/hyaluronidase (Stemcell), mammary epithelial cells were further isolated using 0.25% trypsin (Gibco) and 5 mg/mL dispase (Merck). GFP+ mKate2+ shRNA-expressing luminal and basal cells were sorted via FACS using EpCAM and CD49f surface markers (antibodies 12-9326-42 from eBioscience and 313631 from Biolegend). RNA was extracted using Rneasy Plus Micro kit (Qiagen).  The whole amount of RNA from each sample was used for library prep. RNA-Seq libraries were generated using the NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB #E7770/E7775) according to the manufacturer's protocol. 14 cycles were used for library amplfification.