GSM5835450: HD180_iPSC_2; Homo sapiens; RNA-Seq
iBMECs were subcultured at the end of the differentiaiton on six-well plates coated with 50 μg/mL human placental collagen IV and 25 μg/mL fibronectin from human plasma in endothelial media supplemented with 1% penicillin-streptomycin and 10 μM ROCK inhibitor Y27632.  After one hour, media was switched for an additional 24 hours in the same media, before switching to human endothelial serum free media supplemented with 1% penicillin-streptomycin and 10 mM ROCK inhibitor after another 24 hours. At day 2 of subculture (for iBMECs) or day 3 of culture for iPSCs, cells were washed with 1x PBS and lysed using RLT buffer supplemented with β-mercaptoethanol, before isolating RNA using a RNeasy Mini Kit (Qiagen) with DNase I digestion (Qiagen). RNA libraries were prepared for sequencing using standard Illumina protocols conducted by Novogene.