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. Author manuscript; available in PMC: 2010 Aug 28.
Published in final edited form as: Biochem Biophys Res Commun. 2009 Jun 16;386(3):477–482. doi: 10.1016/j.bbrc.2009.06.059

Fig. 3.

Fig. 3

Chondrogenic and osteogenic differentiation of sorted GFP-positive and negative cells. (A–E) Chondrogenic differentiation. (A) Alcian blue staining of GFP-positive and negative cells. GFP-positive cells showed the most intense staining after 8 days of BMP2 treatment at 500 ng/ml. (B–E) Quantitative PCR for chondrogenic markers after BMP2 treatment. BMP2-treated GFP-positive cells showed highest Col2a1 (B) and Aggrecan (C) expression after 8 days of BMP2 treatment at 500 ng/ml. (D,E) Time course of Col2a1 (D) and Col10a1 (E) expression in GFP-positive cells cultured in the presence or absence of 250 ng/ml BMP2. RNA was extracted at days 5, 8 and 11. Col2a1 expression decreased, while Col10a1 expression increased between days 5–11 in BMP2-treated GFP-positive cells. (F–H) Osteogenic differentiation. (F) Alkaline phosphatase staining showing strong alkaline phosphatase activity in GFP-positive cells after 4 weeks of osteogenic treatment. (G) Alizarin red staining showing massive calcium deposition in cells with 4 weeks of osteogenic treatment. (H) Semi-quantitative PCR for osteoblastic markers in sorted GFP-positive cells with or without osteogenic induction. (Osx) Osterix; (Bsp) Bone sialoprotein; (Alp) Alkaline phosphatase.