Smad-mediated BMP signalling is decreased in chondrocytes of heat-shocked, dnBMPR zebrafish embryos. (A-D′) Condensing ceratohyal mesenchyme of heat-shocked dnBMPR zebrafish embryos (D,D′) showed reduced levels of p-Smad1/5/9 immunoreactivity at 48 hpf (i.e. 24 after 20-min heat shock), compared with non heat-shocked wild-type (A,A′) and dnBMPR (B,B′) embryos and heat-shocked wild-type embryos (C,C′). (E) Quantitative image analyses (n=3 for each group) revealed significant decreases in p-Smad1/5/9 levels in nuclei of condensing ceratohyal mesenchyme of heat-shocked dnBMPR zebrafish, compared with each control group (*P<0.05, one-way ANOVA and paired Student's t-test). (F-I′) Chondrocytes of heat-shocked dnBMPR zebrafish embryos (I,I′) showed reduced levels of p-Smad1/5/9 immunoreactivity at 72 hpf (i.e. 48 h after 20-min heat shock), compared with control groups (F-H′). (J) Quantitative image analyses (n=3 for each group) revealed significant decreases in p-Smad1/5/9 levels in nuclei, but not in non-nuclear regions, of chondrocytes of heat-shocked dnBMPR zebrafish at 72 hpf, compared with each control group (*P<0.05, one-way ANOVA and paired Student's t-test). Scale bars: 50 μm. a.u., arbitrary units; HS, heat-shocked; wt, wild type.
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