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Fig. 2. Adipocyte differentiation of Runx2–/– chondrocytes during culture. Chondrocytes isolated from the ribs of wild-type (D, F) and Runx2–/– (A-C,E,F) embryos at E18.5 were inoculated at a density of 1x105/plate in 24-well plates, and cultured for up to 9 days. (A) Immunocytochemistry using anti-type II collagen antibody of Runx2–/– chondrocytes 1 day after plating. (B,C) Runx2–/– chondrocytes were stained with Alcian blue (B) or with Alcian blue and oil red O (C) at 6 days after plating. Cells with vacuoles are surrounded by proteoglycan, as shown by staining with Alcian blue (B, arrows). (D-F) Wild-type (D, F) and Runx2–/– (E,F) chondrocytes were stained with oil red O 9 days after plating. Low-magnification views are shown in (F). The plates were also stained with hematoxylin and eosin (H-E) to confirm the presence of the cells (F). Magnification A-E, x50.





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