Journal: PLoS ONE

Article Title: Saccharomyces boulardii Improves Intestinal Epithelial Cell Restitution by Inhibiting ?v?5 Integrin Activation State

doi: 10.1371/journal.pone.0045047

Figure Lengend Snippet: αvβ5 integrin is required for Sb S-stimulated cell restitution. (A) Polarized HCT-8/E11 cell monolayers were pretreated without or with 10 µg/ml of an anti- αv integrin mAb (anti-αv) for 1 h. Cell monolayers were then wounded as described in Methods and incubated for 5 h in the absence (ctrl) or presence of Sb S in a medium either containing anti-αv integrin mAb or not. Phase contrast images were acquired at the indicated times. Data shown are from a representative experiment out of 3 performed. Scale bar: 300 µm. (B) HCT-8/E11 cell monolayers were wounded and incubated with Sb S. The monolayers were further incubated without (-) or with the following function-blocking anti-mAbs 1 h before wounding and during cell migration: αvintegrin (αv), αvβ3 integrin (αvβ3), αvβ5 integrin (αvβ5), or αvβ6 integrin (αvβ6). Wound closure was determined as described in Methods . Results are expressed as the percentage of cell migration compared to Sb S-treated cells without mAbs. Data represent the mean+SD of 5 separate experiments. Rat-anti DPP IV (DPPIV) and mouse anti-α5 subunit (α5) were used as isotypic Abs controls. *** P

Article Snippet: To assess the role of the integrins in Sb S enhanced enterocyte migration, 10 µg/ml of the following function-blocking anti-integrin mAbs were used: αv integrin (69.6.5, Beckman Coulter), αvβ3integrin (mouse IgG1, LM 609 clone, Millipore), αvβ5integrin (mouse IgG1, P1F6 clone, Millipore), or αvβ6 (mouse IgG2a, 10D5 clone, Millipore).

Techniques: Incubation, Blocking Assay, Migration

Journal: Virology Journal

Article Title: Integrins are not essential for entry of coxsackievirus A9 into SW480 human colon adenocarcinoma cells

doi: 10.1186/s12985-016-0619-y

Figure Lengend Snippet: CV-A9 infection in the SW480 cell line is independent of an RGD and αV-integrins. a Fluorescence from flow cytometric analysis of integrin expression. SW480 cells were incubated with integrin specific monoclonal antibodies against β1, αVβ3 or αVβ6 (cell controls as shown in a white plot were incubated with a secondary antibody) and 10,000 cells were measured in each sample. The data was analyzed by the Cyflogic program and the area of positive signal is indicated in a white plot. In the case of αVβ3 the white area is shadowed by the negative dark background. Standard threshold values were used in the assays. b The ratio (SW480/A549) of integrin mRNA levels measured by quantitative PCR is indicated. c Immunofluorescence images of SW480 cells infected with the wild type CV-A9 and with the RGD-deletion mutant (CV-A9RGDdel). Antibodies specific to CV-A9 ( green ) were used. The nuclei ( blue ) were stained with Hoechst. The scale bar = 20 μm

Article Snippet: The function-blocking antibodies were against integrin αV (L230; ATCC), integrin αVβ3 (MAB1976Z; Chemicon®), integrin αVβ5 (MAB1961Z; Chemicon®), integrin αVβ6 (MAB2077Z; Chemicon®), integrin β1 (MAB2253; Chemicon®) and integrin α5β1 (MAB1969; Chemicon®).

Techniques: Infection, Fluorescence, Flow Cytometry, Expressing, Incubation, Real-time Polymerase Chain Reaction, Immunofluorescence, Mutagenesis, Staining