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Fig. S6
Metronidazole treatment of tg(osx:NTRmcherry) transgenics leads to efficient osteogenic cell ablation and impaired bone formation. (A) Red channel of images shown in Fig. 4G,J, with dorsal views an calvarial plates of osx:NTRmcherry transgenic fish after treatment with DMSO (left panel) or MTZ (right panel), revealing ablation of approximately 95% of all osx transgenic / osteogenic cells. (B) qRT-PCR analysis of isolated calvarial plates from SL11-12 tg(osx:NTRmcherry) fish. Expression of osteoblast marker genes bglap, col10a1 and col1a1 is strongly reduced after osteogenic cell ablation / MTZ-treatment, compared to the DMSO-treated controls. (C,D) in situ hybridization, revealing loss of col1a1 expression along calvarial plates after treatment with DMSO and MTZ (D), compared to DMSO treatment alone (D). (E,F) Magnified dorsal view of central head region of SL9-10 non-transgenic sibling control fish treated with DMSO (E-E′′; n=5/5), and tg(osx:NTRmcherry) transgenic treated with DMSO + MTZ (F-F′′; n=6/6) for 7 days, after consecutive in vivo alizarin red staining (red; E,F) before and calcein staining (green; E′,F′) after the treatment. Panels (E′′,F′′) show merged images. After osteogenic cell ablation, growth at the osteogenic fronts of the frontal calvarial plates ceases completely (compare with Fig. 1D-F for the strongly reduced, but not completely ceased calvarial growth upon RA-treatment of wild-type fish).