Inhibition of BMP signalling by early DMH1 treatment reduces perichondral bone formation in fam20b mutants down to wild-type levels at 10 dpf. (A-H″) Compared with DMSO-treated wild-type controls (A-B″), perichondral bone appeared to decrease in DMH1-treated wild types at 10 dpf (i.e. 6 days after end of 48 h treatment; E-F″). Similarly, perichondral bone at 10 dpf appeared to decrease in DMH1-treated fam20b mutants (G-H″), compared with DMSO-treated fam20b mutants (C-D″), with levels similar to that seen in DMSO-treated wild types (A-B″). (I) Quantitation of 20 embryos for each experimental group confirmed significant decreases (*P<0.05, one-way ANOVA and paired Student's t-test) in perichondral bone of DMH1-treated wild-type or fam20b−/− embryos. Remarkably, the levels of perichondral bone in DMH1-treated fam20b mutants was statistically indistinguishable from DMSO-treated wild-type controls. Scale bars: 200 μm (A-H). A, anterior; AB/AR, Alcian Blue/Alizarin Red; Ch, ceratohyal cartilage; Chb, ceratohyal bone; Hm, hyomandibular bone; Hs, hyosymplectic cartilage; HS, heat-shocked; L, lateral; n.s., not significant; wt, wild type.
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