Fig. 3.

Itaconate deficiency leads to increased lipid content in foam cells. A) Oil red O staining of peritoneal foam cells (FCs) from Acod1^+/+ → Ldlr^−/− (left) and Acod1^−/− → Ldlr^−/− (right) mice, B) quantified by calculating lipid area per nucleus stained with hematoxylin (n = 5/4 for Acod1^+/+ → Ldlr^−/−/Acod1^−/− → Ldlr^−/−, unpaired t-test). C) Volcano plot of lipidomics from FCs (red = increased in Acod1^−/− → Ldlr^−/−; blue = decreased in Acod1^−/− → Ldlr^−/−; grey = not significant). Dotted line represents significance cut-off (p < 0.05). D) Total lipid content measured in all lipid species from lipidomics (n = 5/4, unpaired t-test). E) Lipid species enrichment analysis from the lipidomics dataset. F) qPCR-derived gene expression of proteins involved in lipid transport and G) their accompanying transcription factors from FCs (n = 5/4, unpaired t-test). H) One of the three aortic root valve cusps in Acod1^+/+ → Ldlr^−/− (left) and Acod1^−/− → Ldlr^−/− (right) transplanted mice stained with toluidine blue where an area of foam cells (red) was measured and nuclei were counted to I) calculate cell size (area/nuclei count; n = 17/22, unpaired t-test). Data points display individual mice and error bars show SD. FC = Foam Cell; AUC = Area Under the Curve; NES = Normalized Enrichment Score. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)