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. 1999 Jan 11;144(1):161–173. doi: 10.1083/jcb.144.1.161

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Gene expression of CDMP-1 transgenic mice. (A) Northern blot analysis using total RNAs extracted from limb buds of normal (lane 1), 742- CDMP1 (lane 2), and 742-CDMP1-Int (lane 3) transgenic mice at 14.5 d.p.c. For each lane, 20 μg RNA was loaded, transferred to the nylon membrane, and hybridized with corresponding probes indicated at the left of the panels. The bottom panel shows the ethidium bromide–stained gel before transfer. Transgenic CDMP-1 mRNA was expressed strongly compared with endogenous Gdf5/CDMP1 mRNA (arrowhead). The expression levels of Ihh and Col10a1 mRNAs, markers for chondrocyte differentiation, were elevated in the transgenic mice. (B) Distribution of CDMP1 mRNA in the forelimb of normal (left), 742-CDMP1 transgenic (middle), and 742-CDMP1-Int transgenic (right) mice at 14.5 d.p.c. (Top) Staining with hematoxylin and eosin. (Bottom) In situ hybridization of semiserial sections using CDMP1 antisense probe. In the transgenic mice, CDMP-1 mRNA was distributed in proliferating chondrocytes, but not in hypertrophic chondrocytes. Note that the height of the hypertrophic zone was increased in transgenic mice. (C) CDMP1 and Col11a2 expression in the rib of normal (left) and 742- CDMP1-Int transgenic (right) mice at 16.5 d.p.c. (Top) Staining with hematoxylin and eosin. (Middle) In situ hybridization of CDMP1 antisense probe of semiserial sections. (Bottom) Col11a2 antisense probe. In the transgenic mice, CDMP-1 mRNA was distributed in proliferating chondrocytes, but not in hypertrophic chondrocytes. The localization of CDMP-1 mRNA was identical to that of Col11a2 mRNA. (D) Immunohistochemical analysis of the primordial cartilage of the tarsals of normal (left) and 742- CDMP1-Int transgenic (right) mice at 14.5 d.p.c. (Top) Staining with hematoxylin and eosin. (Bottom) Immunostaining of anti–CDMP-1 polyclonal antibody showed transgenic cartilage contained a significant amount of CDMP-1. r, resting chondrocytes; p, proliferating chondrocytes; h, hypertrophic chondrocytes. Scale bars, B, 500 μm; C, 200 μm; D, 50 μm.