Fig. S2

Fig. S2

Gene expression in the cartilage callus and adult jaw. (A-D) Colorimetric RNA in situ hybridization shows gene expression in the cartilage callus. The expression of sp7 is not yet visible in the mesenchyme at 6 dpr and evident in the cartilage callus by 10 dpr. At 14 dpr, ctgfa is expressed in a subset of chondrocytes within the callus. Also at 14 dpr, ihhb is expressed weakly in some repair chondrocytes; inset shows stronger expression in the remnant growth plate of the ceratohyal cartilage. (E) Three-color fluorescent in situ hybridization shows co-expression of col1a1a (red), col2a1a (green), and col10a1 (blue) within repair chondrocytes at 10 dpr. Note the similar level of expression of col1a1a in the bone (top right). Nuclei are detected with Hoechst (white). (F) Transgenic col2a1a^BAC:GFP fish have distinct and non-overlapping expression of GFP in Meckel′s cartilage and col1a1a in periosteum at 2 mpf. (G) A subset of Sp7+ cells (green, detected by anti-Sp7 antibody) also express an osteocalcin:GFP transgene (red, detected by anti-GFP antibody). Inset corresponds to boxed region and shows both Sp7-only cells and Sp7+/osteocalcin:GFP+ cells. Note nuclear Sp7 localization compared to cytoplasmic and nuclear GFP. (H) A subset of sp7:GFP+ cells express col1a1a. Scale bars = 100 microns.