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. Overall design: 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.