Three-week-old male LDLR–/– mice were fed a low-cholesterol diet (LCD, 0.02% cholesterol) or high-cholesterol diet (HCD, 0.5% cholesterol) for 8 weeks before mating with female LDLR–/– mice. The F1 offspring were weaned at 3 weeks old and were fed an LCD for 16 weeks. (A) Schematic representation of experimental design and generation of F1 offspring. (B) Serum total cholesterol and triglyceride levels were measured (n = 4–6, ***P < 0.001, 2-sample, 2-tailed Student’s t test). (C) Lipoprotein fractions (VLDL-C, LDL-C, and HDL-C) were isolated from serum, and the cholesterol levels of each fraction were measured (n = 5–6, **P < 0.01, ***P < 0.001; 2-sample, 2-tailed Student’s t test). (D) Quantitative analysis of the lesion area in the aortic root of LCD- and HCD-fed LDLR–/– mice (n = 7, ***P < 0.05, 2-sample, 2-tailed Student’s t test). Representative images are shown to the right. VLDL-C, very low-density lipoprotein cholesterol; LDL-C, low density lipoprotein cholesterol; HDL-C, high density lipoprotein cholesterol. All data are plotted as mean ± SEM. Scale bar: 100 μm.