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Fig. 1. Western blot analysis of PC expression in HGEC. Total protein was extracted from cells cultured either on plastic (control) or on increasing concentrations of collagen IV (A,D), laminin (B,E), or GBM (C,F) substrate. Equal amounts of total protein per substrate concentration (40 µg for A, 50 µg for B,C) were analyzed on SDS-PAGE and immunoblotted with 3D3 anti-PC antibody diluted 1:20 in TBS-5% milk. Blots were stripped and re-probed with anti-tubulin antibody. Lanes 1-5 (A-C), represent cell culture on substrates (0-20 µg/cm2). Quantification of the protein content of PC was performed by densitometric analysis (D-F). The bars represent the mean±s.d. from three independent experiments after normalizing to the density of ß-tubulin. Results were analyzed using single factor ANOVA and Newman-Keuls test (SNK) (*P<0.05 by ANOVA and SNK test; comparison of each different substrate concentration with zero concentration).





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