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Increased superoxide production was observed in all layers of the vascular wall, namely the endothelium, media and adventitia, 14 and may also increase vasoconstrictor tone via stimulation of the expression of endothelin-1 in the smooth muscle and endothelial cell layer

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Immunoprecipitation and immunoblotting Extraction of proteins from cultured cells was performed with a modified lysis buffer (50 mM Tris-HCl [pH 7.5], 0.1% SDS, 1% Triton X-100, 150 mM NaCl, 1 mM dithiothreitol, 0.5 mM EDTA, 100 M PMSF, 100 M leupeptin)