Figure 2

Figure 2. FO prevents PDS-induced bone resorption in zebrafish larvae. Zebrafish larvae (n = 20) at 5 days post fertilization (dpf) were pretreated with 20 ?M PDS for two days prior to treatment with FO (0?100 ?g/mL) for another two days. Fresh media with FO and/or PDS were replenished at 7 dpf. (A) At 9 dpf, zebrafish larvae were stained with 0.03% calcine and observed under fluorescence microscopy. Numbers show vertebrae (right panels). (B) Each vertebral number was manually counted and indicated. (C) Relative bone area and (D) bone intensity were calculated using imageJ software and expressed. All data are presented as means ± standard error of the mean (* p < 0.05 and *** p < 0.001). In a parallel experiment, total mRNA was extracted, and RT-PCR was performed to evaluate the gene expression of (E) osteoblast-related marker genes such as zRUNX2a, zRUNX2b, zOSX, and zALP and (F) osteoclast-related marker genes such as zCTSK, zNFATc-1, zRANK, and zACP5b. z?-Actin was used as the internal control. All data are presented as means ± standard error of the mean (^### p < 0.001 vs. untreated zebrafish larvae; ** p < 0.01, and *** p < 0.001 vs. PDS-treated zebrafish larvae). z: zebrafish; RUNX2a/b: runt-related transcription factor 2a/b; OSX: osterix; ALP: alkaline phosphatase; CTSK: cathepsin K; NFATc-1: nuclear factor of activated T-cells; cytoplasmic 1; RANK: receptor activator of nuclear factor ?B; and ACP5b: Acid phosphatase 5b.