Journal: bioRxiv

Article Title: Rotary jet-spun porous microfibers as scaffolds for stem cells delivery to central nervous system injury

doi: 10.1101/239194

Figure Lengend Snippet: MSC compatibility with PRM scaffolds. (A) Characterization of PRM scaffolds by scanning electron microscopy. (B) BrdU and phalloidin immunostaining of MSCs on coverslips or PRM scaffolds. Cells were able to adhere and incorporate BrdU into DNA (proliferating cells) in either condition (n = 2, experiment in triplicates). Scale bar = 100 μm. (C) After seven days in culture, MTT assay showed similar survival of MSCs when cells were cultured on coverslips or PRM scaffolds (p > 0.05, Student’s t test, n = 5). (D) TUNEL assay showed that MSCs cultured on coverslips or PRM scaffolds did not undergo apoptosis (n = 2, experiment in triplicates). Scale bar = 100 μm. (E) MSCs cultured on coverslips or PRM scaffolds were induced to differentiate in osteocytes and adipocytes. After 3 weeks, cells were stained with Oil Red (staining of lipid vesicles in red) or Alizarin Red (staining of calcium deposits in red). MSCs cultured on PRM scaffolds preserved multipotency (n = 2, experiment in triplicates). Upper scale bar = 50 μm; lower scale bar = 200 μm. (F) PRM induced a 50% increase in CXCL12 secretion by MSCs ( p < 0.05, Student’s t test, n = 5). MSCs: mesenchymal stem cells, PRM: polymeric rough microfibers, BrdU: 5-bromo-2’-deoxyuridine, MTT: 3-(4,5-dimethylthiazol-yl)2,5-diphenyltetrazolium bromide, TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling, DAPI: 4’,6-diamidino-2-phenylindole.

Article Snippet: CXCL 12 in conditioned media was quantified using Mouse CXCL12 DuoSet ELISA (R&D systems, Minneapolis, USA).

Techniques: Electron Microscopy, Immunostaining, MTT Assay, Cell Culture, TUNEL Assay, Staining, End Labeling

Journal: Breast Cancer Research : BCR

Article Title: Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

doi: 10.1186/bcr3053

Figure Lengend Snippet: Genes differentially regulated by IRF5 in MDA-MB-231 cells.

Article Snippet: Briefly, 100 ng/ml human recombinant CXCL12/SDF-1 (R&D Systems, Minneapolis, MN, USA) was added to 600 μl of phenol red-free DMEM medium supplemented with 10% FBS in the lower chamber.

Techniques: Expressing

Journal: Breast Cancer Research : BCR

Article Title: Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

doi: 10.1186/bcr3053

Figure Lengend Snippet: IRF5 reduces CXCR4 cell surface expression and SDF-1/CXCL12-dependent chemotaxis of MDA-MB-231 cells . A . CXCR4 expression (grey line) in unstimulated cells, shown superimposed on the isotype control (grey shaded area), and CXCR4 expression (black line) after stimulation, was measured by flow cytometry. MDA-MB-231 cells (pBabe and pBIRF5) were treated with the CXCR4 ligand SDF-1 for six hours and CXCR4 expression measured. IRF5 expressing cells show no significant expression of CXCR4. M1, Marker 1. Representative histogram plots from three independent experiments performed in duplicate are shown. B . Cells overexpressing IRF5 are incapable of SDF-1-induced migration when compared to empty vector (EV pBabe) control cells. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference in number of cells migrated between pBabe and pBIRF5 cells; * denotes P < 0.02, ** P < 0.005. C . CXCR4 promoter reporter activity was analyzed by Dual Luciferase assay. MDA-231-pBabe and MDA-231-pBIRF5 were transfected with pGL3 empty vector or pGL3 CXCR4 5'Δ3 promoter and mock-treated with PBS or 100 ng/ml CXCL12. Data are expressed as the mean relative stimulation ± SD from three independent experiments performed in triplicate. Statistical significance was determined by comparing the difference in promoter activity between pBabe and pBIRF5 expressing cells; * denotes P < 0.05.

Article Snippet: Briefly, 100 ng/ml human recombinant CXCL12/SDF-1 (R&D Systems, Minneapolis, MN, USA) was added to 600 μl of phenol red-free DMEM medium supplemented with 10% FBS in the lower chamber.

Techniques: Expressing, Chemotaxis Assay, Control, Flow Cytometry, Marker, Migration, Plasmid Preparation, Activity Assay, Luciferase, Transfection