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Figure 5. Positive effects of Hh-Ag treatment on chondrogenesis. A,C: Sox9 expression was strongly up-regulated in the Hh-Ag-treated blastema (C) compared with that in the control (A). B,D: Alcian blue staining for differentiated cartilage revealed an increase in cartilage differentiation (arrowheads in D). A-D: Serial sections from the same froglet limb blastema. F-M: Sagittal sections of the limb regenerate at 21 days after amputation were stained with H&E and Alcian blue (F-H,J-L) and with MF20 antibody (I,M). E-M: Sections were taken at various levels: proximal (F,J), middle (G,K), and distal (H,I,L,M) through the regenerate as indicated in E. F,J: Single cartilage and muscle tissue were observed in both control (F) and Hh-Ag-treated froglet stumps (J). G,H,K,L: Branched cartilage was detected in the Hh-Ag-treated regenerate (K,L), whereas single cartilage was formed along the proximal-distal (PD) axis in the control (G,H). I,M: There were few MF20-positive cells in the regenerate. A-D: The anterior side is on the left of all panels and lines indicate the amputation plane. F-M: The anterior side is on the left and the ventral side is in the upper part of all panels. Scale bars = 200 mu m. |
