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Fig. 6. 3D architecture plays a key role in stimulating fibronectin matrix assembly. (A) CHO(B2){alpha}5 cells in medium supplemented with rat fibronectin (FN) were plated on 3D matrix or on 2D fibronectin-coated substrates with no other additions (3D, 2D) or with conditioned medium (+CM) or resuspended matrix added (+resus matrix). 3D matrix was incubated with rat fibronectin but without cells (3D, right lane). DOC-insoluble material was prepared after 16 hours and fibronectin was detected with IC3 antibody. (B) Confluent WI-38(VA13) cells were lysed and incubated with a 2D fibronectin substrate. CHO(B2){alpha}5 cells were plated on 3D, 2D and 2D+lysate substrates. DOC-insoluble fractions were prepared and analyzed as in A. (C) CHO(B2){alpha}5 cells were plated on 3D fibronectin matrix (3D), on compressed matrix (Comp) or on 2D substrate in the presence of 5 µg/ml rat pFN. After incubation for 7 hours and 18 hours, DOC-insoluble fractions were prepared and analyzed. The immunoblot was developed with ECL plus western blotting detection system and band intensities were quantified. Numbers below each lane represent relative amounts of fibronectin normalized to 3D samples, which were set to 1 for each time point.





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