Fig. 1

Evaluation of mineralization in alendronate-treated zebrafish. (A) The gross morphology of zebrafish aged at 7 dpf which have been treated with different concentrations of alendronate (10, 20, and 30 µM, left bright-filed panel) from 3 dpf onwards. Calcein staining on control and alendronate-treated embryos at 7 dpf (right green fluorescent panel); (B) Quantification of mineralization degree by detecting the fluorescence intensity at the area of centrum form ring in the notochord. (values are mean ± SD; tested by one-way ANOVA pairwise comparison; N = fish number; Different letters indicate significant differences) (C) Chemical structure of alendronate.

Fig. 2

Evaluation of mineralization of the vertebrate column in dorsomorphin-treated zebrafish. (A) The gross morphology of zebrafish aged at 7 dpf which have been treated with different concentrations of dorsomorphin (10, 20, and 30 µM, left bright-filed panel) from 3 dpf onwards. Calcein staining on control and dorsomorphin-treated embryos aged at 7 dpf (right green fluorescent panel; (B) Quantification of mineralization degree detecting the fluorescence intensity at the area of centrum form ring in the notochord. (values are mean ± SD; tested by one-way ANOVA pairwise comparison; N = fish number; Different letters indicate significant differences) (C) Chemical structure of dorsomorphin.

Fig. 4

Increase of mineralization in pentamidine-treated zebrafish. (A) The gross morphology of zebrafish aged at 7 dpf which have been treated with different concentrations of pentamidine (10, 20, and 30 µM, left bright-filed panel) from 3 dpf onwards. Calcein staining on control and pentamidine-treated embryos at 7 dpf (right green fluorescent panel); (B) Quantification of mineralization degree detecting the fluorescence intensity at the area of centrum form ring in the notochord. (values are mean ± SD; tested by one-way ANOVA pairwise comparison; N = fish number; Different letters indicate significant differences) (C) Chemical structure of pentamidine.

Fig. 5

Increase of mineralization in BML-267-treated zebrafish. (A) The gross morphology of zebrafish aged at 7 dpf which have been treated with different concentrations of BML-267 (10, 20, and 30 µM, left bright-filed panel) from 3 dpf onwards. Calcein staining on control and BML-267-treated embryos at 7 dpf (right green fluorescent panel); (B) Quantification of mineralization degree detecting the fluorescence intensity at the area of centrum form ring in the notochord. (values are mean ± SD; tested by one-way ANOVA pairwise comparison; N = fish number; Different letters indicate significant differences) (C) Chemical structure of BML-267.

Fig. 6

Decrease of mineralization in BML-267ester-treated zebrafish. (A) The gross morphology of zebrafish aged at 7 dpf which have been treated with different concentrations of BML-267 ester (10, 20, and 30 µM, left bright-filed panel) from 3 dpf onwards. Calcein staining on control and BML-267 ester -treated embryos at 7 dpf (right green fluorescent panel); (B) Quantification of mineralization degree detecting the fluorescence intensity at the area of centrum form ring in the notochord. (values are mean ± SD; tested by one-way ANOVA pairwise comparison; N = fish number; Different letters indicate significant differences) (C) Chemical structure of BML-267 ester.

Acknowledgments
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