FIG. 2.

Selection of osteogenic cues. MSCs were cultured in complete αMEM with the indicated growth factors, antibodies, and supplements. (A) Cells were lysed and analyzed by western blotting for expression of Runx2 (48 h) and for the bone matrix proteins, bone sialoprotein II (BSP II) and osteopontin (OPN) (21 days). β-Actin was used as loading control. (B) Cells were lysed after 14 days and analyzed with Arsenazo III assay for total calcium content (n=4). Asterisks indicate statistical significance at p<0.05. (C) Cells were fixed after 14 days and stained with Alizarin Red. Representative photographs are shown in the upper panel, and quantification (n=2) is shown in the lower panel. (D) Cells were lysed after 14 days and assayed for alkaline phosphatase (ALP) activity (n=2). Color images available online at www.liebertpub.com/tea