Journal: PLoS ONE

Article Title: Estradiol Increases Mucus Synthesis in Bronchial Epithelial Cells

doi: 10.1371/journal.pone.0100633

Figure Lengend Snippet: 10 −7 M estradiol (black bars) increased FUT-4, -5 and -6 mRNA expression compared with vehicle control (white bars). Data are expressed as A) absolute change in C t value normalized to HPRT1 and B) fold increase in mRNA expression over vehicle control normalized using HPRT1. Values shown are mean ± SEM of 4 different donors performed in 3 biological replicates. Total fucose sugar residues in the cytoplasmic protein fractions of ALI cultures were determined by lectin binding assays. Fucose residues were significantly increased with 10 −7 M estradiol using fucose binding lectins, C-D) AAA, E-F) LTA and G-H) UEA-1. D, F, and H are densitometric quantifications of C, E, and G, respectively. The intensity of all bands in each lane in detecting total fucose residues were quantified using the software program Image J and normalized to total protein loaded per lane in milligrams. Data is expressed as fold increase over vehicle control. Values shown are mean ± SEM of experiments performed with N = 4 donors. * P<0.05, ** P<0.01 compared against vehicle control. Non-parametric t-tests were used in all statistical analyses.

Article Snippet: Membranes were incubated with primary antibodies against ER-α, ER-β, NFATc1, MUC5AC, or (LTA-lotus tetragonolobus asparagus pea: H1601-1, AAA-anguilla anguilla lectin from fresh water eel: H4901-1, and UEA-1-ulex europaeus lectin from gorse: H2201-1) lectins (EY Laboratories Inc., San Mateo, CA) followed by incubation of secondary HRP-conjugated anti-rabbit, anti-mouse and beta-actin antibodies for 1 h at room temperature, and visualized using an enhanced chemiluminescence substrate (Thermo Scientific, Ontario, CAN) and a Chemigenius imaging system (Syngene, Cambridge, UK).

Techniques: Expressing, Control, Binding Assay, Software

Journal:

Article Title: Release of proinflammatory mediators and expression of proinflammatory adhesion molecules by endothelial progenitor cells

doi: 10.1152/ajpheart.00665.2008

Figure Lengend Snippet: A: phase-contrast and fluorescent images of a cord blood-derived “late” endothelial progenitor cell (EPC) colony. Endothelial cells derived from the long-term culture of mononuclear cells (at least 2–4 wk) can form highly proliferative colonies derived from single circulating endothelial cells. Left: phase-contrast image of an EPC colony derived from umbilical cord blood mononuclear cells. Right: confocal microscopy image of an endothelial colony stained with the endothelial surface stain Ulex europaeus lectin (green = fluorescein isothiocyanate label), the nucleus stain 4′,6-diamidino-2-phenylindole (blue) as well as uptake of acetylated low density lipoprotein (red = DiI label). B and C: flow cytometry phenotyping of EPCs using the endothelial surface marker CD31 [platelet endothelial cell adhesion molecule (PECAM)] and the myeloid surface marker CD45. Late EPCs (or colony-forming EPCs) derived from the long-term culture (at least 3–4 wk) of circulating mononuclear cells from adult blood and umbilical cord blood are positive for the endothelial marker CD31 (PECAM) but not for the myeloid marker CD45 while “early” EPCs [or cultured carcinogenic cells (CACs)] derived from the short-term culture (4 days) of adult mononuclear cells also express the myeloid marker CD45. The negative isotype control antibody staining is shown in gray.

Article Snippet: To confirm the endothelial phenotype, adherent cells were incubated with DiI-labeled acetylated low density lipoprotein (LDL; Molecular Probes-Invitrogen, Carlsbad, CA) for 4 h, and, after fixation, they were incubated with the fluorescein isothiocyanate-labeled endothelial surface stain Ulex europeaus lectin 1 (Biomeda, Foster City, CA) for 1 h, as well as the nuclear 4′,6-diamidino-2-phenylindole stain (Molecular Probes-Invitrogen) for 15 min ( 25 ).

Techniques: Derivative Assay, Confocal Microscopy, Staining, Flow Cytometry, Marker, Cell Culture

Journal: PLoS ONE

Article Title: Blood Group Substances as Potential Therapeutic Agents for the Prevention and Treatment of Infection with Noroviruses Proving Novel Binding Patterns in Human Tissues

doi: 10.1371/journal.pone.0089071

Figure Lengend Snippet: (A) Sections were incubated with anti-A (a,b), anti-B (c,d) and biotinylated Ulex lectin (e,f). (B) Sections were incubated with PBS in place of anti-A, anti-B and Ulex . (C) Sections were incubated with GI.1 VLP and then with anti-GI.1 antibody. (D) Sections were incubated with GII.2 VLP and then with anti-GII.2 antibody. (E) Sections were incubated with GII.6 VLP and then with anti-GII.6 antibody. (F) Sections were incubated with GII.6 after incubation with the PGM-(A + H + ), the flying squid liver (B + H + ) and PGM (A − H + ) preparations, respectively, and then with anti-GII.6 antibody. (G) Section from A blood type were incubated with (a) and without (b) GI.1 VLP and then treated with the PGM-(A + H + ) followed by incubation with anti-GI.1 antibody. The section from B blood type was incubated with (c) and without (d) the VLP from GI.1 and then treated with the flying squid liver (B + H + ) followed by incubation with anti-GI.1 antibody. The section from O blood type was incubated with (e) and without (f) the GII.2 VLP and then treated with the PGM (A − H + ) followed by incubation with anti-GII.2 antibody. Immunostaining of all the sections from (A) to (G) was followed by an ABC detection system. See the details in the Text. Magnifications: (a), (c) and (e) in (A) to (F) and (a) to (f) in (G), ×100; (b), (d) and (f) in (A) to (F), ×400.

Article Snippet: Anti-A, anti-B, anti-Le a and anti-Le b mouse monoclonal antibodies were obtained from Ortho Clinical Diagnosis (Rochester, NY) and biotinylated Ulex lectin was from Seikagaku (Tokyo, Japan).

Techniques: Incubation, Immunostaining