The role of hepatic endothelial c-Kit in regulating the homeostasis and pathogenesis of the liver

To investigate the role of hepatic EC-specific endothelial marker—c-Kit in the regulation of vascular homeostasis and liver pathogenesis. We firtly generated endothelial cell-specific conditional c-Kit knock out and their control mice. LSECs isolated from control and EC-specific c-Kit KO mice were subjected to RNA-seq. Second, c-Kit+ LSECs were purified using anti-CD117 magenetic beads and the eluted fraction was further incubated with anti-CD146 magnetic beads to obtain c-Kit- LSECs. c-Kit- LSECs and c-Kit+ LSECs were thereafter compared through transcriptome analysis as well. To further elucidate the function of c-Kit+ LSECs in modulating NASH pathogenesis of the liver challenged by MCD-diet, bulk CD146+ LSECs were purified from MCD mice (4w) subjected with c-Kit+ or c-Kit- LSECs infusion respectively at the last week, and compared by RNA-sequencing. Overall design: Liver sinusoidal endothelial cells were isolated from indicated mice as described in the summary and were subjected to RNA sequencing, in triplicate or quadruplicate, using Illumina HiSeq 3000.