Aortic lipid staining with Sudan IV. A. Aortas were fixed in 4% (w/v) paraformaldehyde overnight and cut open longitudinally. After rinsing in 70% (v/v) ethanol, specimens were stained with Sudan IV solution for 15 min and decolorized in 80% (v/v) ethanol for 20 min until the normal tissue turned white. Neutral resins were used to block specimens. Images were acquired using camera and analyzed with ImageJ. We then calculated the percentage of plaque coverage. Red staining is indicative of lipid deposition. B. Lipid area/total vessel area for five aortas selected from each group. *
P < 0.05 vs. normal controls; #
P < 0.05 vs. diabetic controls; @
P < 0.05 vs. reduced β2GP I (R-β2GP I); and &P < 0.05 vs. β2GP I.