Figure 9. Trehalose administration in mice is atheroprotective.

(a,b) GFP-LC3 mice (ApoE-KO background) were fed western diet for 2 months and administered vehicle or trehalose (2 g kg⁻¹ given five times per week i.p.) in the final 2 weeks of diet (n=4 mice per group). Representative GFP fluorescence (scale bar, 100 μm; a) and quantification of GFP intensity in aortic roots by confocal microscopy (n=4 mice per group; b) is shown. (c–e) TFEB intensity and nuclear localization in aortic roots of the same cohort used in a was detected by immunofluorescence. Shown are (c) representative aortic root TFEB staining (scale bar, 100 μm), (d) average aortic root TFEB intensity; and (e) TFEB-DAPI co-localization. (f) Diagram summarizing experimental protocol and mice cohorts used for in vivo assessment of trehalose in atherosclerosis. Mice were fed a western diet for 2 months while being administered either vehicle, trehalose or sucrose (disaccharides given both i.p. 2 g kg⁻¹ for three times per week and orally 3% ad libitum in drinking water). (g) Serum cholesterol levels at 2 months of western diet (n=7 mice per group). (h) Quantification of atherosclerotic plaque burden by computer image analysis of Oil Red O-stained aortic root sections in the experiment summarized in f (statistical significance of differences was calculated using Mann–Whitney U-test). (i) Serum trehalose levels in wild-type mice (n=4 per group) after administration of trehalose (3 g kg⁻¹) either by i.p. injection or oral gavage at indicated time points. (j) Quantification of atherosclerotic plaque burden by computer image analysis of Oil Red O-stained aortic root sections in mice fed 2 months of western diet while being administered only oral trehalose (statistical significance of differences was calculated using Mann–Whitney U-test). All data are presented as mean±s.e.m. *P<0.05, two-tailed unpaired t-test except h,j.