Quantifying Atherosclerosis in LDLR-/- Mice

To quantify atherosclerosis in LDLR^−/− mice that were placed on HCD (Research Diets Inc., D12108C), aortic roots and aortic arch were embedded in OCT and frozen at −80 °C. Serial cryostat sections (6 µm) were prepared using tissue processor Leica CM3050. For lesion characterizations, the thoracic-abdominal aorta and aortic root were stained for ORO, macrophages (anti-Mac3, BD Pharmingen, 553322, 1:900) T cells (anti-CD4, BD Pharmingen, 553043, 1:90; anti-CD8, Chemicon, CBL1318, 1:100), MHC-positive cells (anti-MHCII, Novus Biologicals), and vascular smooth muscle cells (SM-α-actin, Sigma, F-3777, 1:500)^{73 (link),83 (link)}. The staining area was measured using Image-Pro Plus software, Media Cybernetics, and CD4+ and CD8+ cells were counted manually.