Figure 4.

RIPK1i accelerated macrophage accumulation and foam cell formation. (A–D) Histological analysis of cross-sections from aortic sinus of ApoE^SA/SA mice receiving RIPK1i for 4 weeks. n = 5 mice per group. (A) Representative IHC images of F4/80 in aortic sinus plaque. Scale bar = 100 μm. (B) Quantification of macrophages (F4/80 positive) are represented. (C) Representative IHC images of α-SMA in aortic sinus plaque. Scale bar = 100 μm. (D) Quantification of smooth muscle cells (α-SMA positive) are represented. (E–H) Peritoneal macrophages isolated from male ApoE^SA/SA mice or male C57BL/6J mice were pretreated with RIPK1i (50 ng/ml) or DMSO for 3 h, then stimulated with oxidized LDL (50 μg/ml) for 24 h. (E) Representative images of Oil Red O stained foam cells for male ApoE^SA/SA mice. (F) Percentage of foam cells in ox-LDL induced peritoneal macrophages for male ApoE^SA/SA mice (left, n = 5) and male wild-type mice (right, n = 3). (G) Relative Cd36 mRNA level in ox-LDL induced peritoneal macrophages for male ApoE^SA/SA mice (n = 6). (H) Lipid metabolism-related genes expression levels in ox-LDL induced peritoneal macrophages for male ApoE^SA/SA mice (n = 5–6 per group). All of the data represented as mean ± SEM; ^*P < 0.05, ^**P < 0.01. Statistical analysis: unpaired student's t-test (B,G), non-parametric Mann-Whitney U test (F,H).