Detection of JUP in thrombi.

a) Tissue section of a coronary thrombus stained with anti-JUP mAb 2C9 and labelled goat anti-mouse antibodies (brown) (1, on the left) or with the secondary labelled goat anti-mouse antibody only as a negative control (2, on the right). Nuclei are stained in blue. Immunoreactivity of JUP is detected intra- as well as extracellularly. b) Tissue section of a coronary thrombus, double-stained with anti-JUP mAb 2C9 (red) and the macrophage marker anti-CD68 mAb (black). In addition, an extracellular staining of the fibrin platelet clot was observed. c) Detection of JUP in lysates of coronary thrombi by immunoblotting. A Western blot with recombinant GST-tagged JUP (lane 1, 107 kD), plaque secretome (lane 2), plasma from an ACS patient (lane 3), and lysates of two thrombi (lanes 4 and 5) were immunoblotted with mAb 2C9. d and e) Western blot of macrophages, that differentiated from monocytes isolated from peripheral blood on day 2 (lane 1), day 5 (lane 2), day 7 (lane 3) and day 9 (lane 3), was immunoblotted with mAb 2G9 (which replaced 2C9) (d) and an anti-GAPDH mAb (e) as a loading control.