GSM7159153: 30_m2cafneg; Mus musculus; RNA-Seq
Tumors were collected from 3 10-week-old FVB background MMTV-PyMT mice and then digested with 2 mg/mL Collagenase I and 2 U/mL Dnase. The cells were stained and suject to sorting to isolate each CAF population as described bellow. Sorted cells were directly collected into lyses buffer as instructed by Qiagen RNeasy Micro kit. RNA was further purified following the kit's protocol. icaf: LIVE/DEAD-, CD45/CD31/EPCAM-, PDGFRb+, Ly6C+; m2cafneg: LIVE/DEAD-, CD45/CD31/EPCAM-, PDGFRb+, Ly6C-, CD146-, PDGFRa+, NCAM-; m2cafpos: LIVE/DEAD-, CD45/CD31/EPCAM-, PDGFRb+, Ly6C-, CD146-, PDGFRa+, NCAM+; vCAF: icaf: LIVE/DEAD-, CD45/CD31/EPCAM-, PDGFRb+, Ly6C-, CD146+, PDGFRa- Samples were prepared according to library kit manufacturer's protocol, indexed, pooled, and sequenced on an Illumina NovaSeq 6000.