Journal: Heliyon

Article Title: Determination of critical-sized defect of mandible in a rabbit model: Micro-computed tomography, and histological evaluation

doi: 10.1016/j.heliyon.2023.e18047

Figure Lengend Snippet: Bone morphogenic protein 2 (BMP2) immunostaining results. (A) Immunohistochemical observation of jaw defects at different healing stages (BMP2 immunostaining × 100). (B) BMP2 expression (n = 6, mean ± standard deviation) was consistent in the 4 mm, 5 mm, 6 mm, 8 mm, and 10 mm groups at different cycles. There was no significant difference among groups (p > 0.05). n. s., not significant.

Article Snippet: Nonspecific binding sites were blocked using normal goat serum (Sigma-Aldrich, USA) for 30 min, after which the tissue sections were incubated with rabbit anti-bone morphogenetic protein 2 (BMP2) polyclonal antibody (1:100, bs-1012 R, Bioss, Beijing), anti-collagen type I (Col I) polyclonal antibody (1:100, bs-10423 R, Bioss, Beijing), anti-CD31 polyclonal antibody (1:100, bs-0195 R, Bioss, Beijing), overnight at 4 °C.

Techniques: Immunostaining, Immunohistochemical staining, Expressing, Standard Deviation

Journal: Frontiers in Toxicology

Article Title: Exploring plasticisers-osteoporosis links and mechanisms: a cohort and network toxicology study

doi: 10.3389/ftox.2025.1617663

Figure Lengend Snippet: Validation of Key Targets’ Osteogenic Relevance (A,B) CCK8 assay to determine MEHP concentration. (C) Alkaline phosphatase (ALP) staining of BMSCs after MEHP intervention. (D–F) Decreased expression of type I collagen (COL1), RUNX2, and BMP2 after MEHP intervention. (G) MEHP reduced the protein levels of CTSD and VCP, (H–M) Significant increase in the expression of COL1, BMP2, and RUNX2 genes during osteogenesis of BMSCs. (N, O) MEHP-mediated inhibition of CTSD and VCP proteins impaired the osteogenic capacity of BMSCs. (P) ALP staining after siRNA treatment.

Article Snippet: The membrane was then blocked with 5% skim milk at room temperature for 1 h. Following blocking, it was incubated overnight at 4°C with primary antibodies against BMP2 (bs-0514R, Bioss, China), RUNX2 (BSM-52672R, Bioss, China), GAPDH (10494-1-AP, Proteintech, China), COL1 (14695-1-AP, Proteintech, China), VCP (82463-1-RR, Proteintech, China), and CTSD (21327-1-AP, Proteintech, China).

Techniques: Biomarker Discovery, CCK-8 Assay, Concentration Assay, Staining, Expressing, Inhibition

Journal: Molecular Medicine Reports

Article Title: Dose-dependent inhibitory effects of zoledronic acid on osteoblast viability and function in vitro

doi: 10.3892/mmr.2015.4627

Figure Lengend Snippet: Effect of ZA on the expression of BMP-2 and phosphorylation of the ERK 1/2 and p38 pathways in MC3T3-E1 cells. The MC3T3-E1 cells were incubated with various concentrations of ZA (0–1 µ M) for 7 days. (A) Levels of secreted BMP-2, measured using an ELISA. (B) mRNA expression levels of BMP-2, determined using reverse transcription-quantitative polymerase chain reaction analysis. (C) Protein levels of BMP-2, p38, p-p38, ERK 1/2 and p-ERK 1/2, detected using western blot analysis and normalized to GAPDH. (D) Quantitative analysis of the blots for BMP-2. (E) Relative expression of p-ERK, normalized to ERK, and relative expression of p-p38 normalized to p38. The results are expressed as the mean ± standard error of the mean (n =3 for each group). * P<0.05, ** P<0.01 and *** P<0.001, compared with the 0 µ M group. BMP-2, bone morphogenetic protein 2; ERK 1/2, extracellular signal-regulated kinase 1/2; p-, phosphorylated; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Primary antibodies against the following targets were used: Monoclonal rabbit anti-p38 antibody (1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA; cat. no. 8690), monoclonal rabbit anti-phosphorylated (p)-p38 antibody (1:1,000; Cell Signaling Technology, Inc.; cat. no. 4511), monoclonal rabbit anti-ERK 1/2 antibody (1:1,000; Cell Signaling Technology, Inc.; cat. no. 4695), monoclonal rabbit anti-p ERK 1/2 antibody (1:1,000; Cell Signaling Technology, Inc.; cat. no. 4370), polyclonal rabbit anti-inactive caspase-3 antibody (1:500; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; cat. no. SC-7148), polyclonal rabbit anti-OCN antibody (1:500; Santa Cruz Biotechnology, Inc.; cat. no. SC-30045), polyclonal rabbit anti-active caspase-3 antibody (1:200; Abcam, Cambridge, UK; cat. no. ab2302), monoclonal rabbit anti-ALP antibody (1:20,000; Abcam; cat. no. ab108337), polyclonal rabbit anti-BMP-2 antibody (1:1,000; Abgent Biotech Co., Ltd., Suzhou, China; cat. no. AP13858c), polyclonal rabbit anti-Runx2 antibody (1:400; Wuhan Boster Biological Technology, Ltd.; cat. no. BA3613-2), monoclonal mouse anti-glyceraldehyde phosphate dehydrogenase (GAPDH) antibody (1:400; Wuhan Boster Biological Technology, Ltd.; cat. no. BM1623), monoclonal mouse anti-β actin antibody (1:400; Wuhan Boster Biological Technology, Ltd.; cat. no. BM0627).

Techniques: Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: IP-10/IL-17 co-treatment-initiated HCASMC calcification is mediated by the BMP6 autocrine effect. a The HCASMCs were either maintained as the control or were co-treated with IP-10 and IL-17 for 3 days, after which we examined the BMP2/4/6 mRNA expression in the HCASMCs; b – d the HCASMCs were either maintained as the control or were pre-treated with IgG, BMP2-, BMP4-, or BMP6-blocking antibodies and then co-treated with IP-10 and IL-17; b HCASMC calcification was analyzed using ARS stain; c we examined the mRNA expressions OPN, OCN, and ALP using real-time PCR; d the protein expressions of OPN, OCN, and ALP were examined using the western blot test; e the HCASMCs were either maintained as the control or were treated with IP-10, IL-17, or both, for 48, 72, or 96 h, after which we examined the smad1/5 phosphorylation using the western blot test; f the HCASMCs were pretreated with control-, smad1- or smad5-specific siRNA and then either maintained as the control or co-treated with IP-10 and IL-17 for 14 days. The calcification of the HCASMCs was analyzed using ARS stain. The data in ( a – f ) are mean ± SEM from three independent experiments. * P < 0.05 vs. control cells; # P < 0.05 vs. IgG/or si-CL/IP-10 and IL-17-co-treated cells. The results in ( d – e ) are representative of three independent experiments with similar results

Article Snippet: IP-10, IL-17, and BMP6 ELISA kit, BMP2-specifc rabbit polyclonal antibody, BMP4-specific mouse monoclonal antibody, and BMP6-specific goat polyclonal antibody were come from the Biocompare (San Francisco, CA).

Techniques: Control, Expressing, Blocking Assay, Staining, Real-time Polymerase Chain Reaction, Western Blot, Phospho-proteomics

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: Runx2 regulates the BMP6 autocrine effect on the IP-10/IL-17 co-treatment-initiated OPN/OCN/ALP expression and calcification of HCASMCs. a The HCASMCs were either maintained as the control or were co-treated with IP-10 and IL-17 for 2, 4, 6, and 8 days; b the HCASMCs were either maintained as the control or were pre-treated with IgG or BMP6-blocking antibodies and then co-treated with IP-10 and IL-17 for 8 days; c the HCASMCs were pretreated with smad1- and smad5-specific siRNA and then either maintained as the control or co-treated with IP-10 and IL-17 for 8 days; a – c the expression of runx2 was examined using the western blot test; d – e the HCASMCs were pretreated with control- or runx2-specific siRNA and then maintained as either the control or co-treated with IP-10 and IL-17; d we examined the mRNA expressions of OPN, OCN, and ALP through real-time PCR; e the calcification of the HCASMCs was analyzed using ARS stain. The results in ( a – e ) are representative of three independent experiments with similar results. The data in ( d – e ) are mean ± SEM from three independent experiments. * P < 0.05 vs. siCL/control cells; # P < 0.05 vs. siCL/IP-10 and IL-17-co-treated cells

Article Snippet: IP-10, IL-17, and BMP6 ELISA kit, BMP2-specifc rabbit polyclonal antibody, BMP4-specific mouse monoclonal antibody, and BMP6-specific goat polyclonal antibody were come from the Biocompare (San Francisco, CA).

Techniques: Expressing, Control, Blocking Assay, Western Blot, Real-time Polymerase Chain Reaction, Staining

Journal: Cell & Bioscience

Article Title: Serum IP-10 and IL-17 from Kawasaki disease patients induce calcification-related genes and proteins in human coronary artery smooth muscle cells in vitro

doi: 10.1186/s13578-020-00400-8

Figure Lengend Snippet: KD serum has higher levels of secreted BMP6. The plasma levels of BMP6 from eight non-KD febrile controls and eight KD patients were examined using an ELISA assay. * P < 0.03 vs. febrile controls

Article Snippet: IP-10, IL-17, and BMP6 ELISA kit, BMP2-specifc rabbit polyclonal antibody, BMP4-specific mouse monoclonal antibody, and BMP6-specific goat polyclonal antibody were come from the Biocompare (San Francisco, CA).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay