Fig. 3
Topical 12-O-tetradecanoylphorbol-13-acetate (TPA) application does not affect atherogenesis in apolipoprotein E deficient (ApoE−/−) mice. Plaque areas measured in control and TPA mice in: a cross sections of the aortic root (μm2), and b the aortic arch en face (% of the aortic arch area); data represent mean values, parametric t-test. In a, the number of sections quantified were n = 4–7/mouse in study 1, and n = 1–4/mouse in study 2. c Quantification of the level of macrophages and monocytes (MOMA-2, brown) and collagen content (Trichrome, blue) in aortic root plaque (depicted as % of the total plaque area in the aortic root, mean values, parametric t-tests). Data from study 1 (n = 5–7/group) are shown as follows: control: unfilled circle; TPA: filled circle, and from study 2 (n = 10–15/group) as control: unfilled triangle; TPA: filled triangle. Also shown representative photos of the two aortic root stainings, with scale bar = 200 μm. d mRNA levels of the macrophage marker F4/80, vascular adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), inducible nitric oxide synthase (iNOS), and monocyte chemoattractant protein 1 (MCP-1) in the aortic arch as measured by real-time quantitative PCR. The expression levels were normalized to the housekeeping gene glycealdehyde-3-phosphate-dehydrogenase (GAPDH). Subsequently, fold expression in TPA mice relative to control mice was calculated and depicted (control mice set to 1 and depicted as a dotted line). Ten mice/group from study 2 were randomly selected for this analysis. Results are shown as median (IQR), and statistical differences were analysed with non-parametric t-test