(A) The specificity of the antibody generated against hLDAH was tested in HEK293 cells transfected with hLDAH siRNA and overexpressing flag-hLDAH. (B) 15 μg of THP-1 lysates and 30 μg of human endarterectomy lysates were used to assess hLDAH expression by immunoblotting. The figure displays five representative examples from a total of thirteen endarterectomy specimens analyzed; hLDAH was detected in all the samples tested. (C) Immunoblotting with anti-HSL on 70 μg of protein lysates from mouse RAW 264.7 macrophages, human THP-1 macrophages, HEK 293 cells, and endarterectomy specimens. HEK293 cells transfected with HSL were used as a positive control (C+). (D) Immunoperoxidase staining with anti-hLDAH (left panel, brown color) and anti-CD68 (macrophage marker, middle panel, brown color) in consecutive sections of human endarterectomy specimens. Rabbit IgG was used as negative control (right panel). Bar= 100 μM. (E) Immunoblotting with anti-hLDAH on 7.5 μg of protein lysates from RAW 264.7 macrophages, THP-1 macrophages, human monocytes (HM) and human monocyte-derived macrophages (HMDM).