Fig. 4: Long term exposure to hypercholesterolemia generates monocyte-derived KCs with reduced lipid-loading capacity. | Nature Communications

Fig. 4: Long term exposure to hypercholesterolemia generates monocyte-derived KCs with reduced lipid-loading capacity.

From: Loss of embryonically-derived Kupffer cells during hypercholesterolemia accelerates atherosclerosis development

Fig. 4

A KC numbers before and 4, 8, 13 and 21 days after induction of hypercholesterolemia (0 vs 4 days: ****p < 0.0001 and 0 vs 13 days: *p = 0.04). B Representative flow cytometry of CLEC2 + TIMD4- KCs during prolonged exposure of the mice to hypercholesterolemia. C Frequency of CLEC2 + TIMD4- KCs (0 vs 8 days: **p = 0.0015, 0 vs 13 days: ***p = 0.0001 and 0 vs 21 days: ****p < 0.0001). D EmKCs and MoKCs numbers before and at 4, 8, 13 and 21 days after induction of hypercholesterolemia (Panels A, C and D: n = 9, 8, 5, 4 and 8 mice for experimental points 0, 4, 8,13, and 21 days; 2 independent experiments combined) (1-way ANOVA and multiple comparison test; EmKCs: 0 vs 4 days: ****p < 0.0001; MoKCs: 0 vs 8 days: **p = 0.0013, 0 vs 13 days: ***p = 0.0002 and 0 vs 21 days: ****p < 0.0001). E Representative FACS plots showing TIMD4- cells among CLEC2 + KCs in Ldlr−/− and Ccr2−/− x Ldlr−/− female mice subjected to hypercholesterolemia for 3 weeks. F Fold-change in Csf1 expression (normalized to day 0) in livers of Ldlr−/− mice fed HC diet (n = 5, 4, 4, 4 and 4 mice for day 0, 4, 7, 13 and 21, respectively; 1-way ANOVA and multiple comparison test; 0 vs 4 days: *p = 0.026, 0 vs 8 days: ****p < 0.0001, 0 vs 13 days: *p = 0.043 and 0 vs 21 days: **p = 0.007). G Percentage of KI-67+ cells among EmKCs and MoKCs in HC-fed female Ldlr−/− mice for 3 weeks (***p = 0.0007). H Ly6C+ monocytes and MoDMacs in livers of Ldlr−/− mice fed chow (n = 4) or HC diet (n = 4) for 21 days. (Holm-Sidak multiple comparison test; Chow vs HC: Monocytes **p = 0.002 and MoDMacs **p = 0.006). I Granularity (****p < 0.0001) and (J) lipid content (****p < 0.0001) of EmKCs and MoKCs of mice (n = 9) fed HC diet for 3 weeks. K Lipidomic analysis of sorted EmKCs and MoKCs of mice fed HC diet for 3 weeks (n = 3 mice; Chow vs HC, FC: *p = 0.04 and CE: ***p < 0.001). L Heatmap of scavenger receptors mRNA expression in sorted EmKCs and MoKCs from Ldlr−/− mice after 3 weeks of HC diet (n = 4 mice). M CD36 expression (ΔMFI = CD36 MFI minus MFI of non-stained cells) on EmKCs and on VSIG4− and VSIG4+ MoKCs at 3 weeks of HC diet (n = 4 mice) showing lower expression on VSIG4− MoKCs (**p = 0.009). N FITC MFI (bodipy) for EmKCs and for VSIG4- and VSIG4+ MoKCs after injection of oxidized-LDL labeled with bodipy or not (1-way ANOVA; 2 independent experiments combined; EmKCs vs VSIG4+ MoKCs: *p = 0.031 and EmKCs vs VSIG4− MoKCs: ***p = 0.0008). All data in this figure are presented as mean values ± SEM. Statistical significance has been assessed with a two-sided t-test (or multiple t-tests) unless otherwise stated on the corresponding panel legend. Source data are provided as a Source Data File.

Back to article page