GSM7978162: Mouse vessels, Cldn5 iECKO, rep3; Mus musculus; RNA-Seq
The brain tissue was mechanically and enzymatically digested with Collagenase type II (1 mg/ml, cat. #C6885, Sigma). On ice, the washed microvascular fragments (MVFs) were isolated using sheep anti-rat magnetic Dynabeads® (50 µl/brain, cat. #11035, Invitrogen) bound to rat anti-mouse PECAM1 antibody (10 µl/brain, cat. #553370, BD Biosciences) and a DynaMag 2 magnet (Invitrogen). The washed MVFs were then further dissociated with Collagenase IV (1 mg/ml, cat. #17104-019, Gibco) and TrypLE (cat. #A1217701, Gibco) to remove the magnetic beads and obtain single cell solutions. Libraries were constructed as per manufacurers' recommendation (10x Genomics Gene expression v3.1)