Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: A. Immunostaining of FABP5 protein (brown color) in airway epithelial cells from normal smoker. Bar represent 50 µm. B. Immunostaining of FABP5 protein (brown color) in airway epithelial cells from COPD smoker. Bar represent 50 µm. C. FABP5 protein was quantified by densitometry using the Scion Image software (Scion Corporation, Frederick, MD). Data are representative of 5 different subjects per group.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Immunostaining, Software

Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: A. FABP5 mRNA expression is increased post P. aeruginosa infection, however CS exposure decreases FABP5 mRNA expression and prevents bacteria-induced FABP5 mRNA expression. B. Representative Western blot detection of FABP5 and β-actin proteins shows that FABP5 protein levels increased post Pa infection but CS exposure decreases Pa-induced FABP5 protein levels. Data are representative of 3 independent experiments and are expressed as mean ± SEM.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Expressing, Infection, Bacteria, Western Blot

Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: A. CS exposure increases P. aeruginosa colony forming units (CFU) on FABP5 knocked down primary NHBE cells. B. CS exposure significantly enhances IL-8 secretion in the basolateral supernatant of primary NHBE cell cultures knocked down for FABP5. C. CS exposure decreases β-defensin 2 mRNA expression on FABP5 knocked down primary NHBE cells. FABP5 shRNA − indicates cells that were knocked down for Firefly luciferase and FABP5 shRNA + indicates cells that were knocked down for FABP5. Data are representative of 3 independent experiments and are expressed as mean ± SEM.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Expressing, shRNA, Luciferase

Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: A. CS exposure decreases P. aeruginosa colony forming units (CFU) on primary NHBE cells overexpressing FABP5. B. FABP5 overexpression blocks CS-induced IL-8 secretion in the basolateral supernatant of primary NHBE cell cultures. C. FABP5 overexpression augments β-defensin 2 mRNA expression in primary NHBE cells. FABP5 − indicates cells overexpressing GFP and FABP5 + indicates cells overexpressing FABP5. Data are representative of 3 independent experiments and are expressed as mean ± SEM.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Over Expression, Expressing

Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: A. TLR2 mRNA expression was induced in cells knocked down for FABP5. B. TLR4 mRNA expression was induced in cells knocked down for FABP5. C. TLR2 mRNA expression was decreased in cells overexpressing FABP5. D. TLR2 mRNA expression was decreased in cells overexpressing FABP5. Data are representative of 3 independent experiments and are expressed as mean ± SEM.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: A. FABP5 down regulation significantly decreases PPAR-γ activity in primary NHBE cell cultures. B. FABP5 up regulation significantly increases PPAR-γ activity in primary NHBE cell cultures. FABP5 shRNA − indicates cells that were knocked down for Firefly luciferase. FABP5 shRNA + indicates cells that were knocked down for FABP5. FABP5 − indicates cells overexpressing GFP. FABP5 + indicates cells overexpressing FABP5. Data are representative of 3 independent experiments and are expressed as mean ± SEM.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Activity Assay, shRNA, Luciferase

Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: A. PPAR-γ inhibition by GW9662 increases IL-8 secretion in the basolateral supernatant of primary NHBE cell cultures. B. FABP5 PPAR-γ inhibition by GW9662 dcreases β-defensin 2 mRNA expression in primary NHBE cells. Data are representative of 3 independent experiments and are expressed as mean ± SEM.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Inhibition, Expressing

Journal: PLoS ONE

Article Title: Cigarette Smoke Decreases Airway Epithelial FABP5 Expression and Promotes Pseudomonas aeruginosa Infection

doi: 10.1371/journal.pone.0051784

Figure Lengend Snippet: FABP5 exerts host defense and anti-inflammatory functions against P. aeruginosa bacterial infection indirectly by stimulating PPAR-γ activity. PPAR-γ activity increases β defensin-2 expression thus preventing bacterial growth and inhibits inflammatory cytokine (e.g., IL-8) production.

Article Snippet: After blocking with 1% normal rabbit serum (Vector Laboratories, Burlingame, CA), the slides were incubated with rat anti-human FABP5 primary antibody MAB3077 (R&D Systems, Minneapolis, MN) or a rat IgG control overnight at 4°C, followed by incubation with biotinylated secondary antibody.

Techniques: Infection, Activity Assay, Expressing

Journal: The EMBO Journal

Article Title: The histone methyltransferase Setd8 acts in concert with c-Myc and is required to maintain skin

doi: 10.1038/emboj.2011.421

Figure Lengend Snippet: Embryonic and adult skin is disrupted in the absence of Setd8. ( A ) Visualization of GFP as a reporter for Cre-activity in control (left hand panels) and K14CreSetd8 Δ/Δ (right hand panels) embryos at the indicated embryonic days (E). ( B , C ) Mouse embryos at E18.5 with normal expression of Setd8 (control) ( B ) and deleted Setd8 (K14CreSetd8 Δ/Δ ) in skin ( C ). ( D , E ) Haematoxylin & Eosin staining of skin sections from control ( D ) and K14CreSetd8 Δ/Δ ( E ) littermates at E15.5. ( F – H ) Skin sections of control (upper panels) and K14CreSetd8 Δ/Δ (lower panels) littermates at E14.5 ( F ) and E15.5 ( G , H ) labelled with antibodies to Itga6 (green) and keratin 14 (K14; red) ( F ) or p63 (red) ( G ) or p53 (brown) ( H ). Arrows in ( F ) indicate the loss of an epithelial cell layers starting from E14.5. ( I – L ) Histological sections of adult skin expressing inducible Cre-recombinase under control of the K14 promoter (K14CreER) crossed with wild-type mice (Setd8 wt/wt ) ( I ), or animals heterozygous (Setd8 Δ/wt ) ( K ), homozygous (Setd8 Δ/Δ ) ( J ) for the floxed Setd8 allele, or mice carrying one floxed and one null allele for Setd8 (Setd8 Δ/null ) ( L ). ( M , N ) Labelling for FAS and Fabp5 (green) in the presence (K14CreERSetd8 wt/wt ) (left hand panels) or absence of Setd8 (K14CreERSetd8 Δ/Δ ) (right hand panels). Nuclei are counterstained with DAPI (blue) in ( F , G , M , N ). Arrows in ( I–N ) indicate sebaceous glands and arrowheads ( I – L ) point to IFE. Scale bars: 100 μm ( F – N ).

Article Snippet: Primary antibodies against the following proteins were used: Collagen IV (1:600; AB769, Millipore), FAS (1:100; G-11, Santa Cruz), Itga6 (1:200; cd49f, AbD Serotec), Involucrin (1:800; SY-5, kindly provided by Fiona Watt), Ki67 (1:100; SF6, Vector Laboratories Ltd.), FABP5 (1:100; BAF1476, R&D Systems), p63 (1:100; H-137, Santa Cruz), cleaved Caspase-3 Asp175 (1:100; 9661, Cell Signaling), Phospho-H2A.

Techniques: Activity Assay, Control, Expressing, Staining

Journal: International Journal of Molecular Sciences

Article Title: Isolation and Characterization of Squamous Cell Carcinoma-Derived Stem-like Cells: Role in Tumor Formation

doi: 10.3390/ijms141019540

Figure Lengend Snippet: RAD and NRAD-derived tumor characterization. ( A ) Mitotic Index representing the number of cells undergoing mitosis over total cells were counted in RAD and NRAD-derived tumors ** p < 0.01; ( B ) Survivin and Ki67 expression in RAD and NRAD-derived tumors by immunohistochemistry. Scale bar = 200 μm; ( C ) K10, E-FABP and involucrin expression in RAD and NRAD-derived tumors by immunohistochemistry. Scale bar = 200 μm.

Article Snippet: Membranes containing protein were incubated overnight at 4 °C with the following antibodies: anti-survivin rabbit polyclonal (1:1000; Novus Biologicals, Bloomington, MN, USA), or goat anti-human FABP5 (1:1000; R&D Systems, Inc. Minneapolis, MN, USA), or mouse mAB anti-human involucrin (1:6000; Sigma-Aldrich, St. Louis, MO, USA), or anti-human β 1 -integrin (1:500; Santa Cruz Biotechnology, inc. Santa Cruz, CA, USA) or anti-human β-actin (1:5000; Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Derivative Assay, Expressing, Immunohistochemistry