GSM8524113: Control No4; Mus musculus; RNA-Seq
Isolation of hepatic endothelial cells was performed as described before (PMID: 32916216). Mice were deeply anesthetized with Isolfluran (Baxter) and sacrificed by cervical dislocation. Livers were perfused in situ via the portal vein with a 0.05% collagenase/amino acid/saccharide solution (C2674, Sigma-Aldrich). Livers were dissected and mechanically disrupted. Liver tissue from three mice was pooled and digested at 38 °C in collagenase/Gey's balanced salt solution (G9779, Sigma-Aldrich). Then, the tissue was filtered through a 250 μm mesh. Non-parenchymal cells were separated by a 19.3 % Nycodenz (1002424, Axis-Shield, Alere Technologies) gradient. Subsequently, magnetic-activated cell sorting (MACS) was used with anti-CD146 MicroBeads (ME-9F1, 130-092-007, Miltenyi Biotech) to purify endothelial cells. RNA libraries were prepared for sequencing using standard BGISeq-500 protocols