GSM8833425: sample 1 of liver sinusoidal endothelial cells as a control; Mus musculus; RNA-Seq
After a cell suspension was generated, hepatocytes were eliminated via low-speed centrifugation (50xg), and dead cells and erythrocytes were eliminated via density gradient centrifugation using Percoll PLUS (GE17-5445-02; Cytiva). LSECs were selected using MACS CD146 MicroBeads (132-092-007; Miltenyi Biotec). Primary murine LSECs were exposed to hydrostatic pressure (HP) stimulation. After incubation on a Collagen I-coated plate overnight, the cells were serum-starved for 6 hours and then exposed to periodic HP stimulation for repeated cycles (0.01 Hz) between 101 and 135 kPa (absolute) for 8 hours using our original HPS system. The system was maintained at 37°C in a CO2 incubator (5% CO2). Library preparation was carried out using a TruSeq stranded mRNA Library Prep Kit (Illumina, San Diego, CA) following the manufacturer's instructions. Whole-transcriptome sequencing was performed on an Illumina NovaSeq 6000 sequencer in 101-base single-read mode.