Figure 5: HSCs do not contribute to the generation of newly formed hepatocytes.

(a–g) Determination of possible co-localization of hepatocyte marker HNF4α and LratCre-induced ZsGreen was performed by confocal microscopy in untreated mice (n=4; a), mice receiving MCDE diet for 3 weeks followed by 3 weeks recovery on regular chow (n=4; b), mice receiving DDC diet for 4 weeks followed by 3 weeks recovery (n=3; c), mice undergoing BDL for 2 weeks (n=3; d), 9- to 15-week-old Mdr2ko mice (n=3; e), or 2 weeks after partial hepatectomy (n=3; f). The number of HNF4α-expressing hepatocytes, positive for ZsGreen, was quantified from confocal microscope images (g). (h,i) Representative images of primary hepatocytes isolated from control mice (n=3) and mice that received MCDE diet for 3 weeks (n=4), followed by 3 weeks recovery (h). Positive control (‘+ ctrl’) from AAV8-TBG-Cre-injected mice showing ZsGreen-positive hepatocytes (n=1; h, upper right insert). The rare ZsGreen-positive small-size cells were identified as HSCs by their characteristic fluorescent retinoid-containing lipid droplets (h, lower right inserts). ZsGreen-positive hepatocytes were quantified (i). Scale bar, 100 μm. Data are shown as means±s.d.; NS, non-significant (one-way analysis of variance).