Figure 7. p62 deficiency increases atherosclerotic plaque formation and plaque complexity.

Cohorts of control (p62^+/+/mΦATG5^+/+), p62^−/−, mΦATG5^−/− and mΦATG5^−/−/p62^−/− mice (all on ApoE^−/− background) were fed a Western Diet for 2 months for in vivo assessment of atherosclerosis: (a–c,e) Graphs represent quantification of atherosclerotic plaque burden by computer image analysis of Oil red O-stained aortic root sections from each cohort (representative Oil red O-stained aortic roots are shown right; a,e); by en face analysis at the level of aortic arch (b) and whole aorta (c).(d) Cohorts of ApoE^−/− mice transplanted with bone marrow from control (p62^+/+) and p62^−/− mice were fed a Western Diet for 2 months and atherosclerosis quantified as in (a). (f) Macrophage content in aortic root sections was analyzed using immunofluorescence staining using an antibody against MOMA2; representative MOMA2+ areas are shown (left). (g,h,j,k) Apoptosis and necrotic core areas of aortic root sections were determined by quantification of TUNEL immunofluorescence staining (g,j) and acellular areas respectively (h,k). (i,l) IL-1β abundance in atherosclerotic plaque lysates were determined by ELISA. (m) Graph represents IL-1β concentrations in the serum of atherosclerotic mice cohorts of indicated genotypes (n=3–4 wells pooled from at least total of 9 mice). (n) Graph represents quantification of combined apoptotic and necrotic core areas for all 4 groups. For (f–l,n), the number of mice used is shown below each figure. For all panels, data are presented as mean ± SEM. *P<0.05, **P<0.01; #P<0.05, ##P<0.01 (vs. Control). Scale bar, 0.4 mm (a,e); 100 μm (f).