Figure 5.

Defective lesion regression in diabetic mice is improved by normalizing plasma glucose.

(A) Experimental overview: Ldlr^−/− mice were fed a HCD (0.15%) for 16 weeks to develop atherosclerotic lesions. At this time point a group of mice was sacrificed to determine baseline (pre-regression) lesion characteristics. The remaining mice were divided into 3 groups; Reg, Reg+STZ and Reg+STZ+SGLT2i and placed on chow diet for 6 weeks (n=10-11/group).

(B) Blood glucose and total cholesterol levels at baseline and 6 weeks of regression. *P<0.05 vs. all groups.

(C) Blood leukocyte levels at baseline and after 6 weeks of regression as assessed by flow cytometry. n=9-11/group. *P<0.05 vs. all groups, ^P<0.05 vs. Reg+STZ.

(D) Quantification of mean lesion areas.

(E) Representative ORO stained lesions and quantification of ORO stain as percent of lesion area.

(F) Representative CD68⁺ stained lesions and quantification as CD68⁺ area/lesion. *P<0.05 vs. Reg and Reg+STZ+SGLT2i, ^P<0.05 vs Reg+STZ.

(G-H) FACS isolated Ly6-C^hi monocytes were (G) allowed to adhere to cultured human aortic endothelial cells under static conditions or (H) assessed for their ability to migrate towards CCL2 (in a transwell chamber). *P<0.05 vs. all groups, ^P<0.05 vs. Reg+STZ n=4-6/group.

(I) Representative images and quantification of lesions stained with an anti-Ly6-C antibody (FITC; green) and Hoechst dye (blue; nuclei) using confocal microscopy. Arrows indicate Ly6-C⁺ cells (Ly6-C^hi monocytes). *P<0.05 vs. all groups, ^P<0.05 vs. Reg+STZ. All values are means ± SEM.