Journal:

Article Title: Interleukin-6 Enhances Production of Anti-OspC Immunoglobulin G2b Borreliacidal Antibody

doi: 10.1128/IAI.69.7.4268-4275.2001

Figure Lengend Snippet: Titers of borreliacidal antibody in supernatants obtained from cultures of lymph node cells containing B. burgdorferi isolate 50772 and 1, 10, or 100 μg of anti-IL-6 after incubation for 7 days. Lymph node cells were obtained from C3H/HeJ mice 28 days after vaccination. Control cultures were treated with a goat isotype-nonspecific IgG antibody.

Article Snippet: They were then washed five times with PBS-T and incubated for 45 min at 21°C with 50 μl of biotinylated rat anti-murine IL-6 (MP5-32C11 clone; PharMingen) diluted in PBS-FBS to 1 μg/ml.

Techniques: Incubation

Journal:

Article Title: Interleukin-6 Enhances Production of Anti-OspC Immunoglobulin G2b Borreliacidal Antibody

doi: 10.1128/IAI.69.7.4268-4275.2001

Figure Lengend Snippet: Immunoblots of B. burgdorferi isolate 50772 after treatment with supernatant obtained from cultures of lymph node cells containing B. burgdorferi isolate 50772 and 100 μg of anti-murine IL-6. Supernatant was collected on day 7 of incubation. Lymph node cells were obtained from C3H/HeJ mice 28 or 35 days after vaccination. Control cultures were treated with a goat isotype-nonspecific IgG antibody.

Article Snippet: They were then washed five times with PBS-T and incubated for 45 min at 21°C with 50 μl of biotinylated rat anti-murine IL-6 (MP5-32C11 clone; PharMingen) diluted in PBS-FBS to 1 μg/ml.

Techniques: Western Blot, Incubation

Journal: Journal of Virology

Article Title: Basal Expression Levels of IFNAR and Jak-STAT Components Are Determinants of Cell-Type-Specific Differences in Cardiac Antiviral Responses

doi: 10.1128/jvi.01172-07

Figure Lengend Snippet: FIG. 6. Basal IFNAR expression in cardiac fibroblasts and in cardiac myocytes. Basal levels of IFNAR1 (A and B) and IFNAR2 (C and D) were assessed 2 days postplating in cardiac myocytes (A and C) and cardiac fibroblasts (B and D). Cell suspensions were fixed, permeabilized, and stained with control (solid black line) or with the indicated monoclonal antibody (filled gray histogram). Representative histograms are shown; all samples were stained and run in duplicate.

Article Snippet: The antibodies used were those against an intracellular epitope of IFNAR1 (sc-7391; Santa Cruz Biotechnology) and an extracellular epitope of IFNAR2 (BAF1083; R&D Systems).

Techniques: Expressing, Staining, Control

Journal: Biomedicines

Article Title: Therapeutic Potential of Luteolin on Impaired Wound Healing in Streptozotocin-Induced Rats

doi: 10.3390/biomedicines9070761

Figure Lengend Snippet: Effects of luteolin on protein expression levels of inflammatory factors of skin wound on day 14. ( A ) Tissues were homogenized in RIPA buffer and subjected to immunoblotting with anti-MMP-9, anti-NF-κB, anti-TNF-α, anti-IL-6, and anti-IL1-β antibodies. ( B ) Quantitative results of MMP-9, NF-κB, TNF-α, IL-6, and IL1-β protein levels were adjusted to β-actin protein level. Values of individual groups are presented as mean ± SEM. * p < 0.05, compared with the NDM group. # p < 0.05, compared with the DM group.

Article Snippet: The sections were reacted with primary antibody nuclear factor (NF)-κB (p65) (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA, sc-8008), TNF-α (1:100, Santa Cruz Biotechnology, sc-52746), IL-1β (1:100, Santa Cruz Biotechnology, sc-52012), IL-6 (1:500, Bioss Antibodies, Woburn, MA, USA, bs-0782R-TR), MMP-9 (1:200, Santa Cruz Biotechnology, sc-13520) and vascular endothelial growth factor (VEGF) (1:200, Santa Cruz Biotechnology, sc-7269) at 4 °C for 48 h. After washing with 0.01 M PBS (pH7.4), sections were placed in 1:200 secondary antibody at room temperature for 2 h. Then, the sections were immersed in 1:100 concentration of avidin-biotin complex (ABC complex, Vector) at room temperature for 1 h with 3,3′ diaminobenzidine (DAB, Sigma, St. Louis, MO, USA) as the chromogen.

Techniques: Expressing, Western Blot

Journal: Biomedicines

Article Title: Therapeutic Potential of Luteolin on Impaired Wound Healing in Streptozotocin-Induced Rats

doi: 10.3390/biomedicines9070761

Figure Lengend Snippet: Schematic representation of action by luteolin on impaired wound healing in STZ-induced rats. Luteolin improves diabetic wound healing by inhibiting inflammation and improving antioxidative capacity, which associates with NF-κB-mediates MMP-9, IL-6, TNF-α, and IL-1β downregulation, and Nrf-2-mediates GHS-Px and SOD1 upregulation. Arrows: improve or increase. T-bar: inhibit.

Article Snippet: The sections were reacted with primary antibody nuclear factor (NF)-κB (p65) (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA, sc-8008), TNF-α (1:100, Santa Cruz Biotechnology, sc-52746), IL-1β (1:100, Santa Cruz Biotechnology, sc-52012), IL-6 (1:500, Bioss Antibodies, Woburn, MA, USA, bs-0782R-TR), MMP-9 (1:200, Santa Cruz Biotechnology, sc-13520) and vascular endothelial growth factor (VEGF) (1:200, Santa Cruz Biotechnology, sc-7269) at 4 °C for 48 h. After washing with 0.01 M PBS (pH7.4), sections were placed in 1:200 secondary antibody at room temperature for 2 h. Then, the sections were immersed in 1:100 concentration of avidin-biotin complex (ABC complex, Vector) at room temperature for 1 h with 3,3′ diaminobenzidine (DAB, Sigma, St. Louis, MO, USA) as the chromogen.

Techniques:

Journal: PLoS ONE

Article Title: RNA Sensors Enable Human Mast Cell Anti-Viral Chemokine Production and IFN-Mediated Protection in Response to Antibody-Enhanced Dengue Virus Infection

doi: 10.1371/journal.pone.0034055

Figure Lengend Snippet: (A) CBMCs, HMC-1 and KU812 cells were infected with infectious (DEN+Ab) or UV-inactivated (UV-DEN+Ab) dengue virus in the presence of a subneutralizing concentration of dengue-immune sera (1∶10,000) or were exposed to 10 µg/mL polyI:C (pI:C (10)) or 1 µg/ml polyI:C complexed to Lipofectamine 2000 (pI:C (1)+Lpf). Controls included mock treatment and dengue virus only (DEN). After 24 hours, cell supernatants were harvested and assayed for IFN-α2b and IFN-β by ELISA. Graphs show the mean ± SEM of 4–9 separate experiments. (B) CBMCs, HMC-1 and KU812 cells were treated as in (A). After 12, 24 and 48 hours supernatants were analyzed for IFN-α2b and IFN-β content by ELISA. Graphs show the mean ± SEM of 3–9 separate experiments. (C) CBMCs were infected with dengue virus as described in (A). (C) 48 hour supernatants from CBMCs treated as in (A) were added directly or were blocked for 1 hour on ice with 10 µg/ml sheep anti-human IFN-α2b, goat anti-human IFN-β or appropriate control IgG, prior to addition to fresh KU812 target cells for 24 hours. Cells were then infected with dengue virus with a 1∶10,000 dilution of dengue- immune sera. At 24 hours post-infection KU812 cells were harvested, fixed and permeabilized for detection of intracellular dengue virus E protein by FACS. Data are representative of at least three separate experiments. * P <0.05; ** P <0.01.

Article Snippet: Sheep anti-human IFN-α2b antibody (Endogen) and goat anti-human IFN-β antibody (R&D systems, Minneapolis, MN) were used at 1 or 10 μg/ml for blockade.

Techniques: Infection, Virus, Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Chest

Article Title: Lower Respiratory Tract Myeloid Cells Harbor SARS-Cov-2 and Display an Inflammatory Phenotype

doi: 10.1016/j.chest.2020.10.083

Figure Lengend Snippet: A-E, Lower respiratory tract myeloid cells that harbor SARS-CoV-2 virions display an inflammatory phenotype. A, Quantitative immunofluorescence with median percentage (n=3 slides per patient) of total endotracheal aspirate cells that expressed SARS-CoV-2 nucleocapsid protein (n=6 patients; patient 7 did not have sufficient endotracheal aspirate for immunofluorescence staining). B, Representative montage from a single polymorphonuclear cell shows co-localization by immunofluorescence. Panels from left to right show merge, CD14 (green), IL-6 (red), SARS-CoV-2 nucleocapsid protein (white), and Imaris (Bitplane) surface-rendered image of the overlapping areas of labeling. The blue nuclear stain in all panels is DAPI; the white scale bar is 10 microns. C, Comparison of endotracheal aspirate cells that co-expressed CD14 (14) or CD16 (16) with (N+) or without (N-) SARS-CoV-2 nucleocapsid protein in each sample (n=5 patients; patient 2 was removed due to low number of cells with nucleocapsid protein). Statistical comparison by Mann-Whitney test. The double asterisks indicate a probability value of <.01. D, Comparison of endotracheal aspirate cells that co-expressed IL-6 (IL-6+) or tissue factor (TF+) with (N+) or without (N-) SARS-CoV-2 nucleocapsid protein in each sample (n=5 patients; patient 2 was removed due to low number of cells with nucleocapsid protein). Statistical comparison by Mann-Whitney test; the single asterisk indicates a probability value of <.05; the double asterisks indicate a probability value of <.01. E, Representative in situ localization of CD14 (green), IL6 (white), and tissue factor or F3 (red) transcript and DAPI nuclear staining (blue) in an endotracheal aspirate myeloid cell. Pt = patient; SARS-CoV-2 = severe acute respiratory syndrome coronavirus 2.

Article Snippet: Antibodies that were used include SARS-CoV-2 nucleocapsid protein (NB100-56576; Novus Biologicals), CD14 (#347490; BD Biosciences), CD16 (MA1-84008; Invitrogen), CD142 (Tissue Factor; BD 550252; BD Biosciences), and IL-6 (Novus NBP2-44953; Novus Biologicals).

Techniques: Immunofluorescence, Staining, Labeling, Comparison, MANN-WHITNEY, In Situ