Journal: Molecular Medicine Reports

Article Title: Bone morphogenetic protein-9 promotes the differentiation of mouse spleen macrophages into osteoclasts via the ALK1 receptor and ERK 1/2 pathways in vitro

doi: 10.3892/mmr.2016.5803

Figure Lengend Snippet: Effects of BMP-9 on the expression of BMP receptors. (A) Expression of BMPR-IA, BMPR-IB, BMPR-II and ALK1 receptors in cells treated with BMP-9 (100 ng/ml) using ELISA. **P<0.01 and *P<0.05, vs. other receptors. (B) Immunofluorescence of (a) ALKI receptor, (b) counterstaining with DAPI and (c) the two stains merged in cells without BMP-9 treatment. Immunofluorescence of (d) ALKI receptor (e) counterstaining with DAPI and (f) the two stains merged in cells treated with BMP-9 (100 ng/ml). Positive expression of ALK1 receptor is indicated by white arrows. BMP-9, bone morphogenetic protein-9; BMPR, BMP receptor; ALK1, anaplastic lymphoma kinase 1; OD, optical density.

Article Snippet: The cells were fixed with 4% paraformaldehyde and blocked with 5% BSA, and were then incubated overnight at 4°C with the following primary antibody: Polyclonal goat anti-mouse ALK1 immunoglobulin (Ig) G (1:100; cat. no. 770-MA; R&D Systems, Inc., Minneapolis, MN, USA), quenched with glycine (0.1 M) for 1 h, incubated with secondary antibodies (polyclonal donkey anti-goat IgG; 1:200; cat. no. NL003; R&D Systems, Inc.) for 2 h at room temperature, and then washed three times with PBS.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Molecular Medicine Reports

Article Title: Bone morphogenetic protein-9 promotes the differentiation of mouse spleen macrophages into osteoclasts via the ALK1 receptor and ERK 1/2 pathways in vitro

doi: 10.3892/mmr.2016.5803

Figure Lengend Snippet: Effects of BMP-9 and ALK1 receptor on the cell signal transduction pathway. (A) Western blot of the phosphorylation of ERK1/2 in cells treated with BMP-9 (100 ng/ml) and (B) quantification (*P<0.01, vs. 0 min). (C) Western blot of the phosphorylation of Smad2 in cells treated with BMP-9 (100 ng/ml) and (D) quantification (P>0.01, vs. 0 min). (E) Western blot of the phosphorylation of ERK1/2 in cells pre-treated with siRNA-ALK1 and (F) quantification (*P<0.01, vs. BMP-9). ERK1/2, extracellular signal-regulated kinase 1/2; p-ERK1/2, phosphorylated ERK1/2; Smad 2, small mothers against decapentaplegic 2; p-Smad2, phosphorylated Smad 2; BMP-9, bone morphogenetic protein-9; ALK1, anaplastic lymphoma kinase 1; siRNA, small interfering RNA.

Article Snippet: The cells were fixed with 4% paraformaldehyde and blocked with 5% BSA, and were then incubated overnight at 4°C with the following primary antibody: Polyclonal goat anti-mouse ALK1 immunoglobulin (Ig) G (1:100; cat. no. 770-MA; R&D Systems, Inc., Minneapolis, MN, USA), quenched with glycine (0.1 M) for 1 h, incubated with secondary antibodies (polyclonal donkey anti-goat IgG; 1:200; cat. no. NL003; R&D Systems, Inc.) for 2 h at room temperature, and then washed three times with PBS.

Techniques: Transduction, Western Blot, Phospho-proteomics, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: Bone morphogenetic protein-9 promotes the differentiation of mouse spleen macrophages into osteoclasts via the ALK1 receptor and ERK 1/2 pathways in vitro

doi: 10.3892/mmr.2016.5803

Figure Lengend Snippet: Effects of the ALK1 receptor and ERK1/2 pathways on BMP-9-induced osteoclast differentiation (A) TRAP staining of cells (a) induced by BMP-9 (100 ng/ml) and RANKL (100 ng/ml), and (b) pre-transfected with siRNA-ALK1 or (c) cultured with U0126 (1,000 nmol/l) prior to BMP-9+RANKL. (Phase contrast microscope; magnification, ×20). White arrows indicate TRAP-positive cells. (B) Protein expression of CTR, determined using an enzyme-linked immunosorbent assay, in cells treated with BMP-9 (100 ng/ml) and RANKL (100 ng/ml), transfectecd with siRNA-ALK1 or treated with U0126 (1,000 nmol/l). *P<0.01, vs. BMP-9+RANKL group. BMP-9, bone morphogenetic protein-9; ALK1, anaplastic lymphoma kinase 1; RANKL, receptor activator for nuclear factor-κb ligand; CTR, calcitonin receptor; siRNA, small interfering RNA; OD, optical density; TRAP, tartrate-resistant acid phosphatase.

Article Snippet: The cells were fixed with 4% paraformaldehyde and blocked with 5% BSA, and were then incubated overnight at 4°C with the following primary antibody: Polyclonal goat anti-mouse ALK1 immunoglobulin (Ig) G (1:100; cat. no. 770-MA; R&D Systems, Inc., Minneapolis, MN, USA), quenched with glycine (0.1 M) for 1 h, incubated with secondary antibodies (polyclonal donkey anti-goat IgG; 1:200; cat. no. NL003; R&D Systems, Inc.) for 2 h at room temperature, and then washed three times with PBS.

Techniques: Staining, Transfection, Cell Culture, Microscopy, Expressing, Enzyme-linked Immunosorbent Assay, Small Interfering RNA

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Inflammatory angiomyolipoma of the liver: an unusual case suggesting relationship to IgG4-related pseudotumor

doi:

Figure Lengend Snippet: Immunohistochemical findings in hepatic inflammatory angiomyolipoma. A. Large mono-, bi- and multinucleated histiocytoid myoid cells strongly expressed HMB45 (Note dendritic features of the larger cells on the right). B. Another area with several slender dendritic spindled cells with over all reticulum-like morphology highlighted by HMB45 immunostaining (note prominent sinusoidal vessels). C. several plasma cells expressed IgG4. D. IgG4-positive plasma cells formed dense aggregates at the interphase between tumor (upper left) and liver tissue (lower right).

Article Snippet: Immunohistochemistry was performed on 3 μ sections cut from paraffin blocks using a Ventana automated system (Ventana Medical Systems Inc., Tucson, AZ, USA) according to the manufacturer’s instructions and the following antibodies: hepatocyte paraffin-1 (HepPar-1), pankeratin (KL-1), CK7, desmin, alpha-smooth muscle actin (ASMA), HMB45, Melan A, protein S100 (polyclonal), CD3, CD15, CD20, CD21, CD23, CD30, CD68, ALK-1, CD1a, CD117, Kappa/lambda light chains, rabbit polyclonal antibody against human IgG (1:2000, Dako) and mouse monoclonal antibody against human IgG4 (clone MCA2098G, 1:100, SeroTec, UK).

Techniques: Immunohistochemical staining, Immunostaining, Clinical Proteomics