Figure 3.

Heterogeneity of ECs in mice liver

(A) t-SNE visualization of re-clustered hepatic ECs.

(B) Vwf^hi and Vwf^low ECs in the periportal area in chow and HFD mice liver. PV, portal vein. BD, bile duct. Arrowheads indicated Vwf^hi ECs, and arrows indicated Vwf^low ECs. Scale bar, 50 μm.

(C) Vwf^hi and Vwf^low ECs in the pericentral area in chow and HFD mice liver. CV: Central vein. Arrowheads indicate Vwf^hi ECs, and arrows indicated Vwf^low ECs. Scale bar, 50 μm.

(D) The percentage of each cluster in hepatic ECs in chow and HFD group.

(E) Detection of Egr1⁺ ECs (ECs-3) in chow and HFD mice liver by immunofluorescence staining. Scale bar, 100 μm.

(F) Detection of Ly6a⁺ ECs (ECs-7) in chow and HFD mice liver by immunofluorescence staining. Scale bar, 100 μm.

(G) The statistics of CD31⁺Egr1⁺ double-positive and CD31⁺Ly6a⁺ double-positive areas in chow and HFD mice liver.

(H) Expression of marker genes for ECs-3 (Egr1, Plk2) and ECs-7 (Ly6a, Ifit1, Ifit3, Isg15) in liver NPCs of chow and HFD group at mRNA level.

(I) Detection of Ly6a^hi ECs (ECs-7) in chow and HFD mice liver by flow cytometry. Ly6a^hi Vegfr2^hi ECs were located on the top right panel of each plot, and their percentages in liver NPCs were shown in red. The statistics were shown on the right. n = 3–5 per group. The data are expressed as mean ± SEM. Statistical significance is shown as ∗ for p < 0.05, ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, and ∗∗∗∗ for p < 0.0001, as evaluated by Student’s t test.