Fig. 6
ECZeb2KO attenuates WD-induced liver damage, steatosis and hypo-vascularization. (a–d) Quantification of liver triglyceride levels (a; n = 2 ECWT/5 ECGFP/6 ECZeb2KO for SD; n = 4 ECWT/5 ECGFP/7 ECZeb2KO for WD), body weight (b; n = 3 ECWT/5 ECGFP/8 ECZeb2KO for SD; n = 4 ECWT/5 ECGFP/7 ECZeb2KO for WD), plasma liver enzyme levels (c: aspartate transaminase (AST, left; n = 2 ECWT/5 ECGFP/7 ECZeb2KO for SD; n = 4 ECWT/5 ECGFP/7 ECZeb2KO for WD) and; d: alanine transaminase (ALT, right; n = 3 ECWT/5 ECGFP/7 ECZeb2KO for SD; n = 4 ECWT/5 ECGFP/7 ECZeb2KO for WD) of mice from the indicated conditions after 24 weeks (w) of standard diet (SD) or western-type diet (WD). (e) Diagram showing quantification of hepatocyte size after 24w of SD or WD (left). The right inset panels show images of liver cross-sections stained with Rhodamine-Phalloidin to label cell contours for all 4 conditions (n = 3 ECWT/5 ECGFP/8 ECZeb2KO for SD; n = 4 ECWT/5 ECGFP/7 ECZeb2KO for WD). Nuclei are counterstained with Hoechst (blue). (f) Quantification of liver vascular branch density (expressed as branch/hepatocyte ratio (top left) or number/area in mm2 (bottom left)) after 24w of SD or WD (n = 3 ECWT/2 ECGFP/7 ECZeb2KO for SD; n = 4 ECWT/5 ECGFP/6 ECZeb2KO for WD). The inset panels on the right show representative images of ENDOGLIN-stained cross-sections (white; top) and corresponding vascular masks (blue; bottom) generated by Q-VAT (Quantitative Vascular Analysis Tool) software for all 4 conditions. Quantitative data are expressed as mean ± s.e.m; *P < 0.05 vs. indicated condition by two-way ANOVA with Tukey post-hoc test. Statistical testing for ALT was performed on the log-transformed data. Pictures in e were taken with an EC Plan-Neofluar 20x/0.50 M27 objective on a Zeiss Axio Imager Z1 equipped with an AxiocamMRc5 and Axiovision software. Pictures in f were taken with a Plan Apo 20x (NA 0.75) objective on a Nikon Eclipse Ni-E with Marzhauser Slide Express 2 equipped with a Hamatsu Orca Flash 4.0 camera and NIS Elements software.
