Fig. 2

EC^Zeb2KO amplifies WD-induced LSEC capillarization early during MASLD. (a) Heatmaps with genes in alphabetical order showing color-coded z-scores of normalized RNAseq data of liver sinusoidal EC (LSEC) signature genes (top) and continuous EC genes (bottom) in LSECs sorted from mouse livers belonging to the indicated condition after 4 weeks (w) of standard (SD) or western-type diet (WD). (b–f) Liver cross-sections of EC^WT (b,d) or EC^Zeb2KO (c,e) mice after 8w of SD (b,c) or WD (d,e) stained for collagen (coll.)-type IV (red) and corresponding quantification (f) of collagen type IV expression presented as area % (n = 10 EC^WT/4 EC^GFP/14 EC^Zeb2KO for SD; n = 11 EC^WT/0 EC^GFP/19 EC^Zeb2KO for WD). (g) mRNA expression (presented as log-transformed or ‘log-T’ data) of capillarization marker Cd34 in whole livers from EC^WT/EC^GFP or EC^Zeb2KO mice after 4w (n = 4 EC^WT/14 EC^GFP/14 EC^Zeb2KO for SD; n = 0 EC^WT/13 EC^GFP/18 EC^Zeb2KO for WD), 8w (n = 5 EC^WT/3 EC^GFP/11 EC^Zeb2KO for SD; n = 11 EC^WT/0 EC^GFP/18 EC^Zeb2KO for WD) and 24w (n = 3 EC^WT/5 EC^GFP/8 EC^Zeb2KO for SD; n = 4 EC^WT/4 EC^GFP/7 EC^Zeb2KO for WD) of SD or WD. (h–l) Representative transmission electron microscopy (TEM) images showing fenestrae (indicated by yellow arrowheads) in sinusoids of EC^WT (h,j) or EC^Zeb2KO (i,k) mice after 8w of SD (h,i) or WD (j,k) and corresponding quantification of the average number of fenestrae (l, left), size of fenestrae (l, middle) and porosity (l, right; n = 1 EC^WT/4 EC^GFP/2 EC^Zeb2KO for SD; n = 1 EC^WT/2 EC^GFP/4 EC^Zeb2KO for WD). SpD: Space of Disse; L: lumen. Quantitative data are expressed as mean ± s.e.m. *: P < 0.05 vs. indicated condition by two-way ANOVA with Tukey post-hoc test. Pictures in (b–e) were taken with an EC Plan-Neofluar 10x/0.30 M27 objective on a Zeiss Axio Imager Z1 equipped with an AxiocamMRc5 and Axiovision software. Pictures in (h–k) were taken with a JEOL JEM1400 microscope.