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. 2012 Dec 6;2:935. doi: 10.1038/srep00935

Figure 3. Phosphorylation of Smad family by H1/fBMP-2 polyhedra and estimation of activity of H1/fBMP-2 polyhedra.

Figure 3

(A) ATDC5 cells were co-incubated with H1/fBMP-2 polyhedra and empty polyhedra (CP-H polyhedra). Phosphorylation of Smad1/5 of ATDC5 cells was assayed by use of phospho-Smad1/5 monoclonal antibody. (B) Activity of BMP-2 from H1/fBMP-2 polyhedra were estimated by comparison with that of rhBMP-2. This activity was quantified by the amount of alcian blue staining of glycosaminoglycan which shows early phase differentiation of cartilage formation of ATDC5 cells. (C) Release of BMP-2 from H1/fBMP-2 polyhedra. H1/fBMP-2 polyhedra were spotted on the membrane of cell culture inserts and ATDC5 cells were seeded in each well of the cell culture plate. The late phase osteogenic differentiation of ATDC5 cells was assayed by ALPase activity.