¹F3 and C-terminal modules enhance supportive activity of fibronectin constructs.

<p>Fibronectin (FN) or recombinant fibronectin constructs, 20 nM, were coated on surfaces of coverslips or 24-well tissue culture plates. After 1 hr incubation at 37°C of fibronectin-null cells on the above proteins, 9 µg/ml FITC-fibronectin was added to culture medium, and cells were incubated for an additional 3 hr. (A) Cells were assessed by fluorescence microscopy as described for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0004113#pone-0004113-g002" target="_blank">Fig. 2</a>. Bar = 20 µm. (B) Assembly of FITC-fibronectin was assayed by immunoblotting after SDS-PAGE of material left after 5 min extraction with 1% deoxycholate (DOC insoluble FN). Anti-FITC and anti-beta actin polyclonal antibody were used for detecting deposited labeled fibronectin and loading control, respectively. The amount of DOC-insoluble fibronectin was normalized with actin in the total extract using duplicate samples and the relative amount is shown as bar graph. Each bar represents mean with standard error, 3 sets of experiments with duplicates in each (n = 6).