Osteoprotegerin Antibody - #DF6824

Figure S1.| The results of western-blot analysis showed that different concentrations of recombinant IGFBP7 increased the protein expression of OPG in osteoblastic cell line MC3T3-E1, whereas the expression of RANKL was not affected after 3 days’ IGFBP7 treatment. OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor-κB ligand.

Figure 4. |Y1R antagonist treatment promoted bone formation and inhibit bone resorption in OVX rats. (A) The Alizarin Red S (yellow arrow head) and TRAP staining (red arrow head) of bone tissues in groups. (B) Immunohistochemical analysis of bone tissue among groups for MMP9 and RUNX2 (x 400). (C, D) Western blotting results of MMP, RUNX2, Cath-K, OPG and RANKL expression in bone marrow from rat femurs. The data are expressed as the means ± SD (n = 6 in each group). ***P<0.001; ****P<0.0001 vs. SHAM, and #P<0.05; ####P<0.0001 vs. OVX by one-way ANOVA and Tukey’s post hoc test

Figure 2. MiR-19b-3p inhibits osteoblast differentiation. (A) RT-PCR analysis was used 443 to detect miRNA-19b-3p expression levels in BMSC-derived osteoblasts after 444 miR-19b-3p-overexpressed or knockdown by corresponding lentivirus. (B) ALP activity in 445 BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by 446 corresponding lentivirus. (C) RT-PCR was used to detect EBF2 mRNA expression levels in BMSC-derived osteoblasts after miRNA-19b-3p overexpression or knockdown by 448 corresponding lentivirus. Western blot assay was used to detect (D and E) EBF2, (F-H) 449 RANKL and OPG protein expression levels in BMSC-derived osteoblasts after 450 miRNA-19b-3p overexpression or knockdown by corresponding lentivirus. (I) The ratio of 451 OPG to RANKL in BMSC-derived osteoblasts after miRNA-19b-3p overexpression or 452 knockdown by corresponding lentivirus (J) Alizarin red staining was used to detect cell 453 mineralization at day 21 after differentiation after miRNA-19b-3p overexpression or 454 knockdown at 100×. Data are analyzed using unpaired Student’s t-tests and presented as the 455 mean ± SD from three independent experiments. * p < 0.05; ** p < 0.01 vs. the indicated 456 group.

Fig. 7.| Histological analysis of the inhibitory effect of PF on LPS-induced bone resorption. (A–C) Representative images of HE and TRAP staining, showing the reduced osteolytic legion and TRAP-positive OCs in the PF-treated groups. (D–G) Representative images of IHC staining of RANKL, OPG, OCN and TNF-α. (H) TRAPpositive OCs number. (H) Quantitative analysis of the expression of RANKL, OPG and RANKL/OPG ratio. The data are presented as the mean ± SD (*p < 0.05,**p < 0.01, ***p < 0.001).

Fig. 5. Differential expression patterns of tooth eruption-related genes and proteins between IF cells and MF cells. (A) Non-induced IF cells and MF cells showed different gene expression patterns; Data are mean ± SD of n = 3 replicates, one-way ANOVA, *p＜0.05, **p＜0.01, ***p＜0.001, ****p＜0.0001. (B) Non-induced and induced IF cells and MF cells showed different protein expression patterns. After induction with CLCCM and HERSCM, there were no significant changes of the expression patterns of tooth eruption-related proteins in IF cells and MF cells. (C) The grey value ratios of Western blotting results; Data are mean ± SD of n = 3 replicates, one-way ANOVA followed by Tukey post hoc test, *p＜0.05, **p＜0.01, ***p＜0.001, ****p＜0.0001. (IF1, MF1 represent the cells cultured by α-MEM; IF2, MF2 represent the cells cultured by α-MEM + CLCCM; IF3, MF3 represent the cells cultured by α-MEM+HERSCM).

Figure 4.Deletion of osteopontin (OPN) promoted the differentiation and mineralization of osteoblasts. A) Osteocalcin-positive rate of SaOS-2 cells was detected by flow cytometry. B) The expression of osteocalcin in SaOS-2 cells was observed by immunofluorescence staining, and the fluorescence intensity was exhibited on the right. C) Alizarin Red S staining was performed on SaOS-2 cells to evaluate the mineralization. D) The protein expression of RANKL, M-CSF and OPG was detected by Western blot.