Figure 1: PP hepatocytes labelled by Mfsd2a-CreER.

Schematic figures showing (a) our knock-in strategy for Mfsd2a-CreER allele using CRISPR/Cas9 by homologous recombination and (b) genetic lineage tracing strategy for Mfsd2a⁺ hepatocytes by Cre-LoxP recombination in Mfsd2a⁺ hepatocytes. (c) Whole-mount fluorescence view of the adult liver from 6-week-old Mfsd2a-CreER;Rosa26-RFP mice. Tamoxifen was induced at 2 days before analysis. Scale bar, 1 mm. (d) Immunostaining for RFP, CK19 and HNF4a on liver sections shows Mfsd2a-expressing hepatocytes in PP zone (P). Scale bars, 100 μm. (e) Immunostaining for RFP, PECAM and 4,6-diamidino-2-phenylindole (DAPI) on liver sections shows Mfsd2a-expressing hepatocytes in the PP zone but not PC zone (*). Scale bars, 100 μm. (f) Isolation of RFP⁻ and RFP⁺ cells by flow cytometry followed by quantitative RT–PCR (qRT–PCR) analysis for expression of RFP and Mfsd2a. Expression level of genes in the RFP⁻ cells was set as 1. Error bars are s.e.m. of the mean for all the quantification in this study. (g) Expression of PP and PC genes detected by qRT–PCR. The x axis denotes RFP⁻ (black) and RFP⁺ (red) groups, and the y axis denotes fold induction.. *P<0.05; n=3, two-tailed unpaired t-test. Each immunostaining image is a representative of four individual samples.