GSM2672127: Foxf1fl/+ EC RNAseq; Mus; RNA-Seq
ChIPseq: Formaldehyde cross-linked chromatin was sonicated using Covaris S220 to 200-300 bp and ChIP was performed using SX-8G IP-STAR robot (Diagenode) using anti-FOXF1 antibody (R&D Systems, AF4798, lot # CAHH0316081).; RNAseq: total RNA was extracted from FACS-sorted endothelial cells using the Qiagen QIAquick RNA isolation kit. Chipmentation procedure was used to create ChIP-Seq libraries PMID:    26280331 RNAseq:Total RNA was amplified using the Ovation RNA-Seq System v2 (NuGEN) according to the manufacturer’s protocol. The libraries were prepared with the Nextera XT DNA Sample Preparation kit (Illumina Technologies). 1 ng of cDNA was suspended in Tagment DNA Buffer, and tagmentation (fragmentation and tagging with the adaptors) was performed with the Nextera enzyme (Amplicon Tagment Mix), incubating at 55C for 10 min. NT Buffer was then added to neutralize the samples. RNA-Seq libraries were prepared by PCR with the Nextera PCR Master Mix, and 2 Nextera Indexes (N7XX, and N5XX) according to the following program: one cycle of 72C for 3min, one cycle of 98C for 30s, 12 cycles of 95C for 10s, 55C for 30s, and 72C for 1min, and one cycle of 72C for 5min.  The purified cDNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Illumina HiSeq2500 following the manufacturer's protocol.