Journal: Molecular & Cellular Proteomics : MCP

Article Title: Secretomic Analysis Identifies Alpha-1 Antitrypsin (A1AT) as a Required Protein in Cancer Cell Migration, Invasion, and Pericellular Fibronectin Assembly for Facilitating Lung Colonization of Lung Adenocarcinoma Cells *

doi: 10.1074/mcp.M112.017384

Figure Lengend Snippet: A1AT triggers the assembly of pericellular polyFN on lung adenocarcinoma cells to promote lung colonization. A, CL1-0 and CL1-5 cells were trypsinized and placed in a suspension culture for 2 h, stained with rabbit anti-FN antibodies followed by a GFP-conjugated donkey anti-rabbit IgG secondary antibody, observed under a fluorescent microscope, and detected using flow cytometry. Both IF and flow cytometry was used to demonstrate the different levels of FN1 assembly on the cell surfaces of CL1-0 and CL1-5 cells. The assembly levels of pericellular polyFN of CL1-5 cells were higher than those of CL1-0. The Olympus FB1000 microscope was used, and the fluorescent filter set for 488 excitation at 600× magnification. B, Evaluation of polyFN1 assembly on the cell surface after scrambled siRNA control and A1AT siRNA transfection was performed using IF and flow cytometry. The pericellular polyFN assembly of CL1-5 cells was inhibited when the expression of A1AT was reduced by transfecting siRNA. C, In the nude mice model, a tail vein injection was performed using the CL1-5 cells treated with the scrambled siRNA control and A1AT siRNA or CL1-0 cells, using five mice for each treatment. After 8 weeks, the mice were sacrificed and the tumor nodules of their lungs were measured. A tumor size of 0.5 mm in diameter was counted as one nodule. A significant inhibition of lung experimental metastasis was observed in mice that were injected with A1AT siRNA CL1-5 cells. Three of five lungs are shown in (D). E, Represented tumor sections of these three groups were stained with H&E. F, CL1-5 cells were incubated in suspension for 2 h in 20% FBS media. The cells were subsequently treated with myelin basic protein (MBP) and soluble, truncated dipeptidyl peptidase IV (DPP IV) for 20 min. The five mice in each group were sacrificed after 8 weeks, and the lungs were removed and fixed in a 3.7% formalin fixative. On injection of the soluble, truncated DPP IV-treated CL1-5 cells, lung colonization among the nude mice was completely inhibited. There are three representative lungs shown in each group.

Article Snippet: To predict the relationship between PKCε and A1AT in the regulation of polyFN assembly in lung colonization, Metacore interactome prediction software (GeneGo, Joseph, MI, USA) was used ( C ).

Techniques: Suspension, Staining, Microscopy, Flow Cytometry, Control, Transfection, Expressing, Injection, Inhibition, Incubation

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Cationic antimicrobial peptide, magainin down-regulates secretion of pro-inflammatory cytokines by early placental cytotrophoblasts

doi: 10.1186/s12958-015-0119-8

Figure Lengend Snippet: A snapshot of the top-scored pathway map based on the enrichment distribution sorted by ‘Statistically Significant Maps’ set using Metacore portal. This pathway map was retrieved from the significant pathways involved in inflammatory responses. Experimental data from all files is linked to and visualized on the maps as thermometer like figures ( red , down-regulated signals; blue , no change) to indicate vector component of the cytokine expression in the treatment groups. The cluster of cyto/chemokines which were markedly affected by AMA in the present study are shown in enclosures with broken red lines. B, binding; CM, covalent modification; Cn, competition; CS, complex subunit; IE, influence on expression; +P, phosphorylation; T, transformation; TR, transcription regulation; Z, catalysis. Green denotes positive effect. Red denotes negative effect

Article Snippet: The enrichment analysis and network constructions were achieved using a Metacore bioinformatics platform (GeneGO, St. Joseph, MI, USA) [ , , ].

Techniques: Plasmid Preparation, Expressing, Binding Assay, Modification, Phospho-proteomics, Transformation Assay