Figure 2

Anti-PCSK9 antibody treatment reduces pro-inflammatory macrophage infiltration and downregulates inflammation in APOE*3Leiden CETP mice. (A) Representative immunostaining for Ly6c in frozen aortic root sections and histomorphometric quantification of Ly6c positive cells in aortic plaques (shown as % of area in aortic plaque) (20 x magnification, bar 100 um) (WTD n = 5, PCSK9-mAb1 n = 12). (B) Representative immunofluorescent Mac-3 (red) and α-smooth muscle cell (α-SMA) (green) staining of frozen aortic root sections and quantification of positive cells in aortic plaques (depicted as % of area in atherosclerotic plaque). Nuclei were stained with DAPI (blue) (WTD n = 6, PCSK9-mAb1 n = 5) (20 x magnification, bar 100 um). (C) Real-time PCR analysis of Il-6, Tnfa, Tgfb and Il-1a mRNA expression in spleen-derived MNC of APOE*3Leiden.CETP mice on a WTD and treated with anti-PCSK9 antibody (PCSK9- mAb1, n = 9) or saline (n = 10) every 10 days for 18 weeks. (C) Representative membranes (3 min and 10 min exposure) of mouse serum cytokine array of WTD (n = 4) and PCSK9-mAb1 (n = 4) treated mice. Pixel density was quantitated by ImageJ and results are depicted as fold over WTD. Data are presented as mean ± SEM and fold over control WTD which was set 1. Unpaired Student’s t-test was performed. *p < 0.05, ***p < 0.001 versus control WTD (Western-type diet).