BMP2 Antibody - #AF5163

Fig. 7: Functional and mechanistic insights of AAT-ZCG for DM bone defects in regulating FoxO1/P53 signaling pathways. A Volcano plot showing differentially expressed genes (DEGs) between experimental and control groups. Red dots indicate up-regulated genes, and blue dots indicate down-regulated genes. B Gene Ontology (GO) enrichment analysis of up-regulated genes. C GO enrichment analysis of down-regulated genes. D Heatmap of the expression levels of DEGs across experimental groups, with specific key genes like FoxO1 highlighted. E Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis of DEGs, identifying significant pathways such as P53 signaling, FoxO1 signaling. F Schematic diagram illustrates the proposed mechanism of AAT-ZCG in activating the forkhead box O1 (FoxO1) and P53 pathways. G Western blot was conducted to evaluate the effects of AAT-ZCG on inflammation and osteogenesis through the FoxO1/P53 signaling pathway. H The effect of AAT-ZCG on the expression of FoxO1, n = 3. I The effect of AAT-ZCG on P53 expression, n = 3. J The effect of AAT-ZCG on the expression of pP53, n = 3. K The effect of AAT-ZCG on the expression of MMP13, n = 3. L The effect of AAT-ZCG on the expression of BMP2, n = 3. M) The effect of AAT-ZCG on the expression of Acp5, n = 3. Data are presented as mean ± SD. Figure 7H-M involved three biological replications (n = 3) and analysis of these experiments’ results were performed using one-way ANOVA. ns, no statistical significance. Statistical significance is indicated as follows: P 

Fig. 8: Evaluation of therapeutic effect of AAT-ZCG in vivo. A, B Disintegration of AAT-ZCG in diabetic rats characterized by in vivo imaging of small animals. C CT images of diabetic cranial defect rats at 4 weeks and 8 weeks in different treatment groups. D BV/TV results of each treatment group at 8 weeks. E BS/BV results of each treatment group at 8 weeks. F Histological analysis by hematoxylin and eosin (H&E) staining and massion staining at 8 weeks. G Immunofluorescence staining of Runx2 and BMP2 at 8 weeks. H, I 8 weeks Runx2 and BMP2 immunofluorescence staining relative fluorescence intensity quantification. Blue: DAPI, excitation wavelength (Ex): 405 nm, endow with pseudo-color: RGB (0, 0, 255); Green: BMP2, Ex: 488 nm, endow with pseudo-color: RGB (0, 255, 0); Red: Runx2, Ex: 640 nm, endow with pseudo-color: RGB (255, 0, 0). Data are presented as mean ± SD. Figure 8B involved three biological replications (n = 3), Fig. 8D-E, H-I involved four biological replications (n = 4). Analysis of these experiments’ results were performed using one-way ANOVA. ns, no statistical significance. *P 

Fig. 4. LncMSTRG25 knockdown inhibited osteogenic differentiation of hBMSCs. (A and B) hBMSCs were transfected with siLncMSTRG25-1, siLncMSTRG25-2, and the negative control and induced to differentiate into osteoblasts for 7 d. To evaluate protein expression levels, Western blotting was performed like PAX8, ALP, BMP2, RUNX2, COLL1, OPN, and OCN. (C and D) qPCR was used to detect the relative expression levels of miR-939-5p, LncMSTRG25-1, PAX8, ALP, BMP2, RUNX2, COLL1, OPN, and OCN. (E) hBMSCs were transfected with siLncMSTRG25-1, siLncMSTRG25-2, or a negative control and induced to differentiate into osteoblasts for 14 d. ARS and ALP staining methods were utilized to identify osteoblast differentiation. (F) Immunofluorescence with specific antibodies was used to detect the expression of PAX8, BMP2, and RUNX2 following 7 d of osteoblast differentiation. Data are the mean ± SD of 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Fig. 5 Osteogenic and osteoclastic differentiation of hydrogel. (A, B) ALP staining. (C, D) ARS staining. (E, F) Immunofluorescence staining of BMP-2 and OCN. (G) ALP activity assay. (H, I) Quantitative analysis of ALP and ARS staining. (J, K) Quantitative analysis of BMP-2 and OCN immunofluorescence staining. (L, M) Immunofluorescence staining and quantitative analysis of CTR immunofluorescence staining. (N) TRAP staining of BMDMs-derived osteoclasts. (NS, no significant difference; *, P 

Fig. 6 The ERK and BMP2 signaling pathway is activated by high extracellular Mg2+ in DPSCs during odontogenic differentiation. a ERK phosphorylation was significantly enhanced in DPSCs treated with 1 mM, 5 mM, and 10 mM Mg2+ compared with the 0 mM Mg2+ group, but p38 and JNK phosphorylation amounts were unchanged. b ERK phosphorylation was reduced by 2-APB. c Consistently, the protein levels of BMP2, BMPR1, and phosphorylated Smad1/5/9 were significantly increased in DPSCs exposed to high extracellular Mg2+. d BMP2, BMPR1, and phosphorylated Smad1/5/9 protein amounts were decreased by 2-APB

Fig. 7 Knock-down of LMBR1 promoted the osteogenic and odontogenic differentiation of PDLSCs. A The potential binding site between miR-758-5p and LMBR1 predicted by bioinformatics. B Dual-luciferase reporter assay demonstrated the correlation between miR-758-5p and LMBR1. C qRT-PCR evaluated the downregulation of LMBR1 by siRNAs. D Western blot evaluated the suppression of LMBR1 by siRNAs. E The results of the CCK-8 assay showed cell proliferation after the downregulation of LMBR1. F Relative mRNA levels of ALP, DSPP, OSX, and RUNX2 when LMBR1 was knocked down. G Western blot assay showed the expression of ALP, DSPP, OSX, and RUNX2 after transfection of siRNAs. H Histogram derived from relative protein expression of western blot. I Result of ARS staining. J Quantification of ARS staining. K Result of ALP staining. L Western blot showed the condition of BMP2 and BMP4 when miR-758-5p was overexpressed or inhibited. M Western blot showed the condition of BMP2 and BMP4 when LMBR1 was knocked down. N Protein levels of ALP, DSPP, RUNX2 and OSX in rescue experiment. O Protein levels of BMP2 and BMP4 in rescue experiment. n = 3, ***P