GSM8181362: retinal endothelial cells, EC-Foxc1-KO, postnatal day 6, biological replicate 1; Mus musculus; RNA-Seq
Retinas pooled from 3 - 6 neonates were digested in collagenase Type I solution (2 mg/mL) for 40 minutes at 37°C with gentle agitation. Cells were then filtered through a 70 μm cell strainer and the pellet was resuspended in Buffer 1(PBS, 0.1% BSA, 2 mM EDTA, pH 7.4). The cell suspension was then incubated with magnetic Dynabeads (Invitrogen) pre-coated with CD31 antibody to isolate the endothelial cell population. After several washes with Buffer 1, RNA was extracted from endothelial cells using RNA STAT solution (Tel-Test) followed by phenol-chloroform treatment. Directional mRNA libraries were prepared using Illumina TruSeq mRNA Sample Preparation Kits per the manufacturer's instructions