Fig. 4. Osteogenic induction of dermal fibroblasts. (A) After 1 week of osteogenic induction, positive staining of alkaline phosphatase (ALP) was observed in the induced group (left panel) but not in the non-induced group (right panel). (B) After 4 weeks of osteogenic induction, intensive calcium deposition was detected by Alizarin Red staining (ARS) in the induced group (left panel), and only very weak staining was observed in the non-induced group (right panel). (C) RT-PCR detects ALP mRNA expression after 1 and 2 weeks of induction, and osteocalcin (OCN) mRNA expression after 2 weeks of induction. No expression of ALP or OCN was observed in non-induced cells. Bars, 50 µm (A); 100 µm (B).