Fate and Functional Roles of Prominin 1+ Cells in Liver Injury and Cancer [Quant-seq]

Purpose: The goal of the experiment is to obtain RNAseq profiles of Prom1+ Ductular Reactive Progenitors (DRPs) from mouse model of severe alcoholic liver injury Method: mRNA-seq analysis was performed in FACS sorted Prom1+CD49f+CD45-, Prom1-CD49f+CD45-, Prom1+CD49f+CD45+ cells and presorted CD45- liver cells. Conclusion: We obtained liver specific Prom1+ genes from analysis. The livers from AH mouse liver were digested with collagenase. Hepatocytes were removed by low centrifugation. Non-parachymal cells stained with CD45 magnetic beads to remove CD45+ cells with MACS. Presorted CD45- liver cells were stained with Prom1 antibody and CD49f antibody. The cells were FACS sorted for three different populations: Prom1+CD49f+CD45- (R6), Prom1-CD49f+CD45- (R5) and Prom1+CD49f+CD45+ (R8). Total RNA from R6, R5, R8 and presorted CD45- cells were extracted. The RNA from two mice were pooled for RNA-seq analysis.