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Fig. S14
Shha-driven Smoothened signaling directs progenitor osteoblasts to align against the basal epidermis but does not influence their proliferation. (A, B) Runx2, sp7 and EGFP immunostaining (red, green, and white, respectively) on longitudinal 96 hpa fin sections from individual shha:GFP fish treated at 48 and 72 hpa with DMSO (panel A) or 40 mg/kg BMS-833923 (panel B). Nuclei are blue. White brackets demonstrate the extension of Runx2+ pObs. BMS-833923 treatment causes misalignment with the distally moving basal epidermis, resulting in both increased layered and incompletely extended Runx2+ pObs. White arrowheads indicate the approximate position of transverse “split” sections shown in Fig. 7C and D. (C) Quantification of the number of Runx2+ pOb cells distal to the first Runx2/sp7+ cells. Each data point represents a scored independent section (10 individual rays from three DMSO treated fish and 13 rays from four BMS-833923 treated fish). (D, E) Runx2 (red) and EdU incorporation (four hour treatment, white) on longitudinal 54 hpa fin sections from individual fish treated at 48 with DMSO (panel D) or 50 mg/kg BMS-833923 (panel E). Nuclei are blue. (F) Quantification of Runx2+ pOb proliferation after DMSO or BMS-833923 treatment. Data represents the scoring of EdU-incorporating pObs from 14 individual rays from three DMSO treated fish and 10 rays from five BMS-833923 treated fish. Error bars are one standard deviation. Scale bars in A and B: 20 μm; D and E: 50 μm.