Fig. 7

Fig. 7 Fig. 7. Combined sup-optimal doses of RA and ECS modulating chemicals is sufficient to influence bone development. Zebrafish were treated from 50 to 72 hpf with 60% combined doses of RA + RIMO, DEAB + OLE, or DEAB + WIN and mineralization of the opercle (black arrowhead) and opercle stem (white arrowhead) was assessed at 144 hpf with von Kossa stain (A–H). RA + RIMO showed no effect on mineralization (C,D). Both DEAB + OLE or DEAB + WIN treatments were sufficient to reduce mineralization (E–H). Measurements of the area of staining of the opercle stem support these observations (M, n = 9). The effects of combined doses on col10a1 expression was assessed at 72 hpf following treatment from 36 to 72 hpf (I–L). RA + RIMO showed no effect on col10a1 expression (J). Both DEAB + OLE or DEAB + WIN treatments were sufficient to reduce expression (K,L). Quantification of staining of the opercle supports the observed expression (N, n = 10).