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Fig. S4
osx:NTRmCherry-encoded protein is expressed in osteogenic cells of all differentiation stages. (A,B) Double fluorescence of tg(osx:NTRmCherry) (red; upper row) together with tg(runx2:EGFP), marking osteoblast progenitors (Kague et al., 2012) (A) or with tg(bglap:GFP) (Vanoevelen et al., 2011) (B), marking mature osteoblasts (green, middle row); counterstained with DAPI (white); transverse sections through frontal plate. Osteogenic front (of) is indicated. Yellow arrowheads in the merged images (lower row) point to osx+runx2 double positive close to the osteogenic front (A), and to osx+bglap double positive bone-lining cells more remote from the osteogenic front (B). (C) Double immunofluorescence for tg(osx:NTRmCherry)-driven RFP and the osteogenic cell marker ZNS5; transverse section through the orbital bone of SL22 wild-type fish: in addition to its co-expression with ZNS5 in bone-lining cells, the osx transgene is expressed in isolated cells within the mineralized bone, which most likely are terminally differentiated osteocytes (Franz-Odendaal et al., 2006).