Fig. 3

The lectin domain of CD248 and the N-terminal 70-kDa fragment of fibronectin mediate CD248-fibronectin interaction. A Schematic presentation of CD248 protein structure and fibronectin fragments used in this study. B Protein stain (Coomassie blue stain) and Western blot analysis of rCD248 protein, including rCD248D1 (6 µg for protein stain and 25 ng for western blot), rCD248D3 (6 µg for protein stain and 140 ng for western blot), and rCD248D1-4 (4.8 µg for protein stain and 40 ng for western blot). C, D Solid-phase binding assay. C Various rCD248 proteins (0.1 µM) were applied to fibronectin (10 µg/mL) - or BSA (as control)-coated multiple−well plate for interaction. The level of interaction was detected with antibodies against the c-myc tag on rCD248 proteins. N = 3. ***P < 0.001. D Various fibronectin proteins, including intact fibronectin (10 µg/mL) and equimolar amounts of its various recombinant fragments (70 kDa, 45 kDa, and 30 kDa) were coated onto a multiple-well plate. rCD248D1-4 (0.1 µM) was applied to the well and the interaction was detected by an anti-c-myc antibody. N = 4. *P < 0.05; ***P < 0.001