Galectin 3

Mice were fed a normal mouse chow (B & K Universal Ltd, UK) and harvested at 9 or 16 weeks of age. For the high fat diet cohort, 8 week old mice were fed a western-type diet for 8 weeks and harvested at 16 weeks of age. Atherosclerotic lesion size was assessed in paraffin-embedded aortic root sections stained with Masson-Goldner trichrome (Merck, Germany). The average lesion size was calculated from three sections taken at 100 μm intervals starting from the section showing all three aortic cusps. The infiltration of macrophages into aortic lesions was analysed using anti-Galectin-3 (BD Pharmingen, UK) immunostaining (Supplementary Table 4). Aortic lipid deposition was assessed in fixed aortas stained with Oil red O (Sigma-Aldrich, UK) from mice fed a chow diet for 16 weeks. Aortic roots were visualised and imaged (coolSNAP-pro camera, Roper Scientific, Leica DMRBE microscope and the lesion area and Galectin-3 positive areas were quantified from digitized microscopic images using Image-Pro Plus (Media Cybernetics, USA).