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. 2020 Jul 9;5(13):e137289. doi: 10.1172/jci.insight.137289

Figure 4. Effects of RAGE and DIAPH1 on macrophage properties.

Figure 4

(A and B) Ki67 (A) and TUNEL (B) staining was performed on aorta sections from diabetic donor Ldlr—/— mice and WT, Ager—/—, and Diaph1—/— diabetic recipient mice. Scale bar: 250 μm. N = 5 mice/group. (C) Schematic of bead assay for tracking monocyte/macrophages in atherosclerotic plaques. (D) Representative images and quantification of the beads per lesion area from sections from nontransplanted diabetic Ldlr—/— and diabetic WT, Ager—/—, and Diaph1—/— recipient mice 5 days after aortic transplantation. Scale bars: 250 μm, and inset, 50 μm. In D, no-bead control is shown (N = 10 mice/group). (E) Role of RAGE and DIAPH1 in reverse transendothelial migration of macrophages. BMDMs from WT, Ager—/—, and Diaph1—/— diabetic mice were subjected to reverse transendothelial migration assays at 2, 24, and 48 hours. N = 6 mice/group. (F) Gene expression of selected pro- (Nos2 and Tnfa) and antiinflammatory genes (Arg1 and Il10) in BMDMs retrieved from BMDM-MAEC cocultures after transendothelial migration assay at 48 hours. N = 4 mice/group. Mean ± SEM. Unpaired t test or Mann-Whitney U test (A, B, and F) depending on the normality of the data. Kruskal-Wallis test followed by Dunn’s multiple comparisons test (D). One-way ANOVA with post hoc Tukey’s test (E). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.