GSM4673227: EC62-SLUG - D2; Homo sapiens; RNA-Seq
The monolayer of Human Lung Fibroblasts seeded on top of the gel were first completely removed and the angiogenic EC were extracted from fibrin-gel using serial trypsinization using 5% trypsin. Cells were then released from the cytodex beads using Collagenase IV. Individual EC were separated from the beads using 100µm cell strainer and centrifugation. The EC were then lysed in RNA lysis buffer and incubated at 55ºC with Protease K (Qiagen 19131) for 15 minutes. The RNA was then isolated following the manufacturer's protocol using a Qiagen RNA Mini Kit (Qiagen 74104). RNA integrity (RIN) was measured for all samples using a Bioanalzyer Agilent 2100. All sequencing libraries analyzed were generated from RNA samples with a RIN score ≥ 9. The Illumina TruSeq mRNA stranded protocol was used to obtain poly-A mRNA from all samples. A total of 200ng of isolated mRNA was used to construct RNA-seq libraries, and these were quantified and normalized using the Library Quantification Kit from Kapa Biosystems
GEO Accession
GSM4673227