Extended Data Fig. 4: Isolation and gene expression profiling of hepatocyte-derived peripheral cholangiocytes.
From: De novo formation of the biliary system by TGFβ-mediated hepatocyte transdifferentiation
a, FACS gates for peripheral cholangiocyte (pC; EPCAM+DBA–) and hilar cholangiocyte (hC; EPCAM+DBA+) isolation from Alb-cre+/−Rbpjf/fHnf6f/f and Rbpjf/fHnf6f/f mice. b, qPCR analysis of floxed Rbpj (Rbpjf/f) genomic DNA in hepatocyte-derived pC (HpC) and hC isolated from Alb-cre+/−Rbpjf/fHnf6f/f mice relative to hepatocytes isolated from Rbpjf/fHnf6f/f mice (dashed line; n = 3 each). Data were normalized to a downstream genomic region of Rbpj to control for gene copy number. Data are mean ± s.e.m. c, d, RNA-seq analysis of normal pC (n = 3 mice), HpC (n = 4 mice) and RBPJ- and HNF6-deficient hepatocytes (H; n = 3 mice). c, Heat map of genes reflecting deletion of Rbpj and Hnf6 (also known as Onecut1). Rbpj mRNA is present in this knockout mouse as a truncated transcript that does not produce a functional protein26. d, Heat map of all differentially expressed CYP genes distinguishing genes associated with mature (M), adolescent (A) and immature (I) hepatocyte differentiation or low expression in the liver (L)29. P < 0.05, one-way ANOVA, FDR-corrected; fold change > 3 (c, d, except Rbpj and Notch1–Notch4). Bold genes denote P < 0.05 for HpC versus pC, two-sided Student’s t-test (c, d).
