Fig. 2: Hypercholesterolemia generates cholesterol-loaded foamy KCs.
A Gene Set Enrichment analysis (GSEA) of the reactome_cholesterol_biosynthesis and foamy plaque macrophages gene signatures in KC from chow- and HC-fed (4 days) Ldlr−/− mice using the Phantasus software. The foamy plaque macrophages gene signature was generated from a previously published dataset (GSE116239). B KCs in HC-fed mice exhibit high granularity (SSC-Ahi) and (C) strongly stain for lipids (bodipy; n = 8 chow and n = 7 HC; combined 2 independent experiments; ****p < 0.0001). D Changes in cholesterol content (*p = 0.0115), in cholesteryl-esters (*p = 0.0115) and various CE species in KCs sorted from chow- and HC-fed (4 days) Ldlr−/− mice (n = 3 chow and n = 4 HC; 1 experiment). E Two-photon laser scanning microscopy images of the liver of chow and HC-fed (4 days) Ldlr−/− mice. Numerous lipid droplets (white spots) are detected by Coherent anti-Stokes Raman Spectroscopy (CARS) in TIMD4+ KCs (red; 3D reconstruction by mask rendering on the right images) and in the hepatocytes (autofluorescence in green) of HC-fed mice as compared to the chow condition. Scale bar = 10 μm. F Calculated amount of total cholesterol (**p = 0.006) and of free cholesterol (FC) and cholesteryl-esters (CE) (****p < 0.0001) present in the liver of chow- (n = 6 mice) and HC-fed (4 days; n = 8 mice; 1 experiment) Ldlr−/− mice. G Calculated amount of total cholesterol (****p < 0.0001) and of FC (**p = 0.003) and CE (**p = 0.003) present in the KC pool of chow- (n = 3 mice) and HC-fed (4 days; n = 5 mice; 1 experiment) Ldlr−/− mice. H Heatmap generated from the RNA-Seq depicting the level of expression of scavenger receptors in KCs from chow- and HC-fed (4 days) Ldlr−/− mice. I Changes in the granularity (SSC-A; ***p = 0.0002) and lipid content (bodipy; **p = 0.005) of KCs in mice fed HC diet overnight and injected in vivo either with anti-CD36 blocking antibodies (n = 8) or isotype control antibodies (n = 9) (2 independent experiments combined). All data in this figure are presented as mean values ± SEM. Statistical significance has been assessed with a two-sided t-test unless otherwise stated on the corresponding panel legend. Source data are provided as a Source Data File.
