Extended Data Figure 9. Ductal specific Tet1 conditional deletion impairs duct-mediated liver regeneration.

a, Schematic of the Prom1Cre^ERT2/Rosa^lslZsGreen/Tet1^flx/flx mouse model. b, Representative immunofluorescence analysis (OPN⁺ red, ZsGreen⁺, green) of Prom1^{ΔTet1/ZsGreen} and Prom1^{Tet1WT/ZsGreen} upon tamoxifen treatment and injection of AAV8-TBG p21 (n=2 mice per genotype). Nucleus, Hoechst. Scale bar, 50 μm c, Representative immunofluorescence analysis of livers from Prom1^{Tet1WT/ ZsGreen} mice injected with AAV8-TBG p21 not receiving tamoxifen treatment (n=2 mice per genotype). Scale bar, 100 μm. d, Tet1 expression in EpCAM⁺/ZsGreen⁺ ductal cells isolated by FACS from Prom1^{ΔTet1/ZsGreen} (n=4) or Prom1^Tet/ZsGreen (n=4) livers derived from mice treated for 3-cycles of DDC and collected 12 days after damage. Graph represents the expression of Tet1 for both genotypes expressed as a fold change compared to Prom1^Tet1WT. Student's two tailed t-test statistical analyses were performed. ***, p<0.001. e, Representative pictures of P21 immunohistochemistry analyses. Scale bar, 200 μm. f, Weight curves of mice undergoing AAV8-TBG-p21 injection followed by DDC treatment (mean± 95%CI). g, TET1 ChIP-qPCR analyses on target genes in ZsGreen⁺/EpCAM⁺ ductal cells isolated from Prom1^{Tet1WT/ZsGreen} DDC-treated livers for 5 days. Cells isolated from 3 mice littermates were pooled used for each independent experiment (n=2). ND, not detected. h, Graph represents mean ±SD of mRNA expression of Tet1 and selected target genes (fold change vs WT undamaged) in EpCAM⁺ ductal cells isolated from undamaged (n=2 per genotype) or day 5 DDC-treated livers (n=3 per genotype) derived from Prom1^{Tet1WT/ZsGreen} (grey) or Prom1^{ΔTet1/ZsGreen} (blue) mice. Statistical analysis was performed using Student's two-tailed t-test compared to the Prom1^{Tet1WT/ZsGreen} value at the corresponding time point.