“…Porcine VSMC were transfected with pCI-lacZ or pCI-TIMP-3 (pCI backbone from Promega) with or without USE as described above. TIMP-3 expression was assayed after 48 h in control and plasmidtransfected cultures by Western blotting 20 mg total cell protein using a mouse monoclonal anti-human TIMP-3 primary antibody that detects human and porcine TIMP-3 6 (1:500; TCS Cellworks Ltd, Buckingham, UK) with a goat anti-mouse IgG-horseradish peroxidase secondary antibody (1:1000; Dako, High Wycombe, UK). Lane a, recombinant human TIMP-3 (23 kDa, Sigma); b, untransfected VSMC; c, USE+BR14 (50% (v/v)); d, lacZ plasmid; e, lacZ plasmid+USE+BR14 (50% (v/v)); f, TIMP-3 plasmid; g, TIMP-3 plasmid+USE+BR14 (50% (v/v)).…”