FIGURE 5.

Fibronectin (FN) transit and matrix assembly in a smooth muscle cell (SMC)-fibroblast scaffold coculture. (a) Fibroblasts and SMCs were cultured on opposite sides of a transwell scaffold for 5 days. Conditioned medium was collected from each side and FN was isolated by gelatin-Sepharose pulldown. FN was separated on an 8% polyacrylamide-SDS gel and immunoblotted with antibodies specific to either human FN (top) or rat FN (bottom). (b) BJ human fibroblasts and WKY3M22 rat SMCs were grown to confluence on unaligned and aligned sides of the scaffold, respectively. After 5 days, samples were fixed and stained with R457 anti-fibronectin antiserum to detect total FN (green) or HFN7.1 anti-human FN antibody (red). Representative images are shown. Scale bar = 50 μm