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    Structured Review

    Thermo Fisher 96 well plates
    Proliferation of PBMC from macaques with collagen-induced arthritis (CIA). PBMC (2 × 10 5 cells in 100 μL per well) were seeded in <t>96-well</t> plates. Cells were cultured with or without CII (40 μg/mL) for 72 h, after which they were evaluated in a BrdU assay. The stimulation index value (mean optical density values of CII-stimulated cultures divided by the mean optical density values of medium-only cultures) was calculated for each treatment. (A) Proliferation of PBMC. (B) Proliferation of PBMC against CII. (C) Stimulation index. Data are expressed as box-and-whisker plots ( n = 3 or 4 per group); group means were compared by using Mann–Whitney U tests. *, Value significantly ( P
    96 Well Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Relationships between Cytokine Levels and Disease Parameters during the Development of a Collagen-induced Arthritis Model in Cynomolgus Macaques (Macaca fascicularis)"

    Article Title: Relationships between Cytokine Levels and Disease Parameters during the Development of a Collagen-induced Arthritis Model in Cynomolgus Macaques (Macaca fascicularis)

    Journal: Comparative Medicine

    doi: 10.30802/AALAS-CM-18-000058

    Proliferation of PBMC from macaques with collagen-induced arthritis (CIA). PBMC (2 × 10 5 cells in 100 μL per well) were seeded in 96-well plates. Cells were cultured with or without CII (40 μg/mL) for 72 h, after which they were evaluated in a BrdU assay. The stimulation index value (mean optical density values of CII-stimulated cultures divided by the mean optical density values of medium-only cultures) was calculated for each treatment. (A) Proliferation of PBMC. (B) Proliferation of PBMC against CII. (C) Stimulation index. Data are expressed as box-and-whisker plots ( n = 3 or 4 per group); group means were compared by using Mann–Whitney U tests. *, Value significantly ( P
    Figure Legend Snippet: Proliferation of PBMC from macaques with collagen-induced arthritis (CIA). PBMC (2 × 10 5 cells in 100 μL per well) were seeded in 96-well plates. Cells were cultured with or without CII (40 μg/mL) for 72 h, after which they were evaluated in a BrdU assay. The stimulation index value (mean optical density values of CII-stimulated cultures divided by the mean optical density values of medium-only cultures) was calculated for each treatment. (A) Proliferation of PBMC. (B) Proliferation of PBMC against CII. (C) Stimulation index. Data are expressed as box-and-whisker plots ( n = 3 or 4 per group); group means were compared by using Mann–Whitney U tests. *, Value significantly ( P

    Techniques Used: Cell Culture, BrdU Staining, Whisker Assay

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    Centrifugation:

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    Luciferase:

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    Reporter Assay:

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    Article Snippet: Paragraph title: Dual-luciferase reporter assay ... The cells were plated into 96-well plates for 24 h and then transfected with BCL6 wild-type or mutant 3'UTR vector using Lipofectamine 2000 reagent (Invitrogen).

    Synthesized:

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    Article Snippet: First-strand cDNA was synthesized from total RNA (0.5 μg) using the High Capacity cDNA synthesis kit with RNase inhibitor (Applied Biosystem, Foster City, CA, USA). .. Quantitative PCR (qPCR) was performed using 10 μl of Fast SYBR Green Master Mix (Invitrogen) with 0.2 μM/l of each primer, 5 μl of 20-fold diluted cDNA and nuclease-free water (Gibco, Carlsbad, CA) to a final volume of 20 μl in 96-well plates (Applied Biosystem, Forster City, CA).

    Cytometry:

    Article Title: Type 2 innate lymphoid cells participate in IL-33-stimulated Th2-associated immune response in chronic obstructive pulmonary disease
    Article Snippet: Paragraph title: Flow cytometry, cell sorting and cell culture ... ILC2 cells were then seeded into 96-well plates at 500 cells/well, mixed with gamma-irradiated PBMCs ( ) from three healthy volunteers (2×106 cells/ml), and cultured in the presence of 500 ng/ml IL-2 in IMDM (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Sigma-Aldrich; Merck KGaA) and 10 ml/l penicillin-streptomycin.

    Quantitative RT-PCR:

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    Electrophoresis:

    Article Title: Effect of growth rate on transcriptomic responses to immune stimulation in wild-type, domesticated, and GH-transgenic coho salmon
    Article Snippet: All pairs of primers were validated for specificity by electrophoresis to confirm the expected amplicon size. .. Quantitative PCR (qPCR) was performed using 10 μl of Fast SYBR Green Master Mix (Invitrogen) with 0.2 μM/l of each primer, 5 μl of 20-fold diluted cDNA and nuclease-free water (Gibco, Carlsbad, CA) to a final volume of 20 μl in 96-well plates (Applied Biosystem, Forster City, CA).

    Incubation:

    Article Title: Antifungal Drugs Influence Neutrophil Effector Functions
    Article Snippet: .. Finally, PMN suspensions were placed in 96-well plates (Nunc; Thermo Fischer Scientific) and incubated for 30 min at 37°C. .. After preincubation with antifungals (50 μl containing 2 × 105 PMN per well), PMN were stimulated with 50 μl TM3 containing either lipopolysaccharides from Salmonella enterica serotype Typhimurium (final concentration of 1 μg/ml; Sigma-Aldrich) or zymosan A from Saccharomyces cerevisiae (zymosan; final concentration of 15 μg/ml; Sigma-Aldrich), with unstimulated PMN serving as controls, for 30 min at 37°C to analyze activation and degranulation.

    Article Title: Downregulation of SNCA Expression by Targeted Editing of DNA Methylation: A Potential Strategy for Precision Therapy in PD
    Article Snippet: .. Adherent NPCs in 96-well plates were incubated with 2 μM MitoSOX (excitation/emission 510/580 nm) and 2 μM MitoTracker Green (485/520 nm) (Life Technologies) in high-glucose medium without phenol red for 15 min at 37°C in the dark. ..

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    Article Snippet: 500 μl 500 m m sodium carbonate and 100 μl Folin & Ciocalteau's phenol reagent were added to the supernatant and incubated 30 min at 37 °C. .. Absorbance was measured in 96-well plates at 660 nm (Electro Multiscan Spectrum, Thermo Fisher Scientific) for 10 min. Activity calculation was performed by the following equation: Acticity [ unit ml ] = released   tyrosine   [ μ mol ] × assay [ m L ] enzyme   volume [ ml ] × incunation   time [ min ⁡ ] × measurement   volume [ ml ] (Eq.

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    Amplification:

    Article Title: Effect of growth rate on transcriptomic responses to immune stimulation in wild-type, domesticated, and GH-transgenic coho salmon
    Article Snippet: All pairs of primers were validated for specificity by electrophoresis to confirm the expected amplicon size. .. Quantitative PCR (qPCR) was performed using 10 μl of Fast SYBR Green Master Mix (Invitrogen) with 0.2 μM/l of each primer, 5 μl of 20-fold diluted cDNA and nuclease-free water (Gibco, Carlsbad, CA) to a final volume of 20 μl in 96-well plates (Applied Biosystem, Forster City, CA).

    Activity Assay:

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    Expressing:

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    Article Snippet: .. For analysis of GFP expression by immunoblotting, transfected cells grown in 96-well plates or on scaffolds were lysed at specified time points in RIPA buffer (Thermo Fisher Scientific) containing protease and phosphatase inhibitors. .. Protein concentration was determined by the BCA Protein Quantification Assay Kit (Pierce).

    BIA-KA:

    Article Title: Aligned Nanofibrillar Scaffolds for Controlled Delivery of Modified mRNA
    Article Snippet: For analysis of GFP expression by immunoblotting, transfected cells grown in 96-well plates or on scaffolds were lysed at specified time points in RIPA buffer (Thermo Fisher Scientific) containing protease and phosphatase inhibitors. .. Protein concentration was determined by the BCA Protein Quantification Assay Kit (Pierce).

    Real-time Polymerase Chain Reaction:

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    Resazurin Assay:

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    Transfection:

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    Article Title: Aligned Nanofibrillar Scaffolds for Controlled Delivery of Modified mRNA
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    SYBR Green Assay:

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    Article Snippet: .. Quantitative PCR (qPCR) was performed using 10 μl of Fast SYBR Green Master Mix (Invitrogen) with 0.2 μM/l of each primer, 5 μl of 20-fold diluted cDNA and nuclease-free water (Gibco, Carlsbad, CA) to a final volume of 20 μl in 96-well plates (Applied Biosystem, Forster City, CA). .. The reaction was performed in triplicate using the 75 Fast Real time PCR System (Applied Biosystem) with the reaction conditions: 95 °C/10 min; 40–45 cycles of 95 °C/2 s, 60 °C/15 s, 72 °C/33 s. After qPCR, a melt curve analysis was performed to verify the presence of a single amplicon peak.

    Cell Culture:

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    Article Snippet: MD NPCs were seeded at 3.5 × 104 cells/mm2 and cultured in high-glucose N2B27 medium without phenol red in black 96-well plates (Greiner). .. Adherent NPCs in 96-well plates were incubated with 2 μM MitoSOX (excitation/emission 510/580 nm) and 2 μM MitoTracker Green (485/520 nm) (Life Technologies) in high-glucose medium without phenol red for 15 min at 37°C in the dark.

    Article Title: Relationships between Cytokine Levels and Disease Parameters during the Development of a Collagen-induced Arthritis Model in Cynomolgus Macaques (Macaca fascicularis)
    Article Snippet: PBMC (2 × 105 cells/100 μL per well) were seeded in 96-well plates (Thermo Scientific, Waltham, MA). .. Cells were treated with CII (40 μg/mL) or not and cultured at 37 °C in a humidified 5% CO2 –air atmosphere for 72 h (total volume, 200 μL per well).

    Article Title: Type 2 innate lymphoid cells participate in IL-33-stimulated Th2-associated immune response in chronic obstructive pulmonary disease
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    Multiple Displacement Amplification:

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    Isolation:

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    Article Snippet: .. To promote proliferation, isolated oligodendrocyte progenitor cells (OPCs) were seeded on 96‐well plates in Neurobasal media (Gibco, Grand Island, NY, USA) with PDGF‐AA (20 ng/mL, PeproTech, Rocky Hill, NJ, USA); bFGF (20 ng/mL, PeproTech, Rocky Hill, NJ, USA); 2% B27 (Gibco, Grand Island, NY, USA); and 1% penicillin/streptomycin at a density of 2 × 104 /well. .. After 2 days, cells were switched to Neurobasal media with 40 ng/mL triiodothyronine (T3) (Sigma‐Aldrich, St. Louis, MO) and 2% B27 to promote differentiation for 7 days.

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    RNA Sequencing Assay:

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    Article Snippet: Paragraph title: Real-time quantitative PCR validation of RNA-Seq data ... Quantitative PCR (qPCR) was performed using 10 μl of Fast SYBR Green Master Mix (Invitrogen) with 0.2 μM/l of each primer, 5 μl of 20-fold diluted cDNA and nuclease-free water (Gibco, Carlsbad, CA) to a final volume of 20 μl in 96-well plates (Applied Biosystem, Forster City, CA).

    Fluorescence:

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    Article Snippet: Adherent NPCs in 96-well plates were incubated with 2 μM MitoSOX (excitation/emission 510/580 nm) and 2 μM MitoTracker Green (485/520 nm) (Life Technologies) in high-glucose medium without phenol red for 15 min at 37°C in the dark. .. Fluorescence was detected by sequential readings, and MitoSOX signals were normalized to mitochondrial content (Mitrotracker) and cell number (Hoechst).

    Mutagenesis:

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    Article Snippet: .. The cells were plated into 96-well plates for 24 h and then transfected with BCL6 wild-type or mutant 3'UTR vector using Lipofectamine 2000 reagent (Invitrogen). .. Cells were lysed 48 h after transfection, and luciferase activity was then determined by the Luc-PairTM Duo-Luciferase HS Assay Kit (GeneCopoeia).

    Mitosox Assay:

    Article Title: Downregulation of SNCA Expression by Targeted Editing of DNA Methylation: A Potential Strategy for Precision Therapy in PD
    Article Snippet: The MitoSox assay was used for the detection of mitochondria-associated superoxide levels. .. Adherent NPCs in 96-well plates were incubated with 2 μM MitoSOX (excitation/emission 510/580 nm) and 2 μM MitoTracker Green (485/520 nm) (Life Technologies) in high-glucose medium without phenol red for 15 min at 37°C in the dark.

    Protein Concentration:

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    Flow Cytometry:

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    Labeling:

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    Purification:

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    Chloramphenicol Acetyltransferase Assay:

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    Plasmid Preparation:

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    Software:

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    Enzyme-linked Immunosorbent Assay:

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    Concentration Assay:

    Article Title: Antifungal Drugs Influence Neutrophil Effector Functions
    Article Snippet: Antifungal stock solutions were diluted with TM3 to working solutions, with desired working concentrations being five times higher than the highest final concentration. .. Finally, PMN suspensions were placed in 96-well plates (Nunc; Thermo Fischer Scientific) and incubated for 30 min at 37°C.

    Article Title: A Stemness and EMT Based Gene Expression Signature Identifies Phenotypic Plasticity and is A Predictive but Not Prognostic Biomarker for Breast Cancer
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    High Throughput Screening Assay:

    Article Title: Downregulation of SNCA Expression by Targeted Editing of DNA Methylation: A Potential Strategy for Precision Therapy in PD
    Article Snippet: High-throughput screening plate reader analysis (FLUOstar Omega, BMG) was conducted following a previously described protocol. .. Adherent NPCs in 96-well plates were incubated with 2 μM MitoSOX (excitation/emission 510/580 nm) and 2 μM MitoTracker Green (485/520 nm) (Life Technologies) in high-glucose medium without phenol red for 15 min at 37°C in the dark.

    FACS:

    Article Title: Type 2 innate lymphoid cells participate in IL-33-stimulated Th2-associated immune response in chronic obstructive pulmonary disease
    Article Snippet: Paragraph title: Flow cytometry, cell sorting and cell culture ... ILC2 cells were then seeded into 96-well plates at 500 cells/well, mixed with gamma-irradiated PBMCs ( ) from three healthy volunteers (2×106 cells/ml), and cultured in the presence of 500 ng/ml IL-2 in IMDM (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Sigma-Aldrich; Merck KGaA) and 10 ml/l penicillin-streptomycin.

    Cell Viability Assay:

    Article Title: A Stemness and EMT Based Gene Expression Signature Identifies Phenotypic Plasticity and is A Predictive but Not Prognostic Biomarker for Breast Cancer
    Article Snippet: For cytotoxicity experiments with Lapatinib and Midostaurin MDA-MB-157 cells (3000 cells/well) were seeded in 96-well plates and twenty-four hours after seeding, cells were transfected with the siZEB1 and siSNAI2 at a final concentration of 20 nM, using Lipofectamine 2000TM (Invitrogen, CA) transfection reagent according to the manufacturer's protocol. .. Cell viability was assessed 72 hours after treatment with CellTiter-Glo Luminescent Cell Viability Assay (Promega, Madison, WI, USA) as per the manufacturer's instructions.

    Gradient Centrifugation:

    Article Title: Relationships between Cytokine Levels and Disease Parameters during the Development of a Collagen-induced Arthritis Model in Cynomolgus Macaques (Macaca fascicularis)
    Article Snippet: Gradient centrifugation with Ficoll-Paque (GE Healthcare Life Sciences, Piscataway, NJ) was performed at 400 × g for 30 min. PBMC were washed twice with PBS without calcium chloride or magnesium chloride (DPBS, Wel Gene, Seoul, South Korea) and adjusted to 2 × 106 viable cells/mL in complete endotoxin-free RPMI 1640 medium (GE Healthcare Life Sciences) supplemented with 10% fetal calf serum (Gibco, Grand Island, NY) and 1× antibiotic–antimycotic (Gibco). .. PBMC (2 × 105 cells/100 μL per well) were seeded in 96-well plates (Thermo Scientific, Waltham, MA).

    Fluorescence In Situ Hybridization:

    Article Title: Effect of growth rate on transcriptomic responses to immune stimulation in wild-type, domesticated, and GH-transgenic coho salmon
    Article Snippet: Real-time quantitative PCR validation of RNA-Seq data The same samples used in RNA-Seq (n = 2 pools of 5 fish per growth response group/treatment) were subjected to qPCR validation for a subset of DEGs. .. Quantitative PCR (qPCR) was performed using 10 μl of Fast SYBR Green Master Mix (Invitrogen) with 0.2 μM/l of each primer, 5 μl of 20-fold diluted cDNA and nuclease-free water (Gibco, Carlsbad, CA) to a final volume of 20 μl in 96-well plates (Applied Biosystem, Forster City, CA).

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  • 99
    Thermo Fisher biofilms
    Loss of outer membrane integrity in strain RN102. Bacterial samples were collected from 48-hour-cultured <t>biofilms</t> for TEM analysis. TEM images of the bacterial cells and the cell appendages are shown for strains: (A) BW25113, (B) RN102, (C) BW25113/pNTR-SD, (D) RN102/pNTR-SD, and (E) RN102/pNT3( hldE ). The outer membranes are indicated by arrows. Representative electron-microphotographs of each strain are shown. A 500-nm-long bar is shown in the lower left corner of each eclectron-micrograph. (F) Western blot analysis of supernatants from BW25113 and RN102. Supernatants were harvested by centrifugation from bacterial liquid culture grown for 48 hours under static conditions. Results of Western blot using anti-Crp, anti-DsbA, anti-OmpC, and anti-OmpA antisera are shown. (G) Supernatants from bacterial liquid cultures of BW25113 or RN102 grown for 48 hours under static conditions were serially diluted with TE. The diluted samples were used as template DNA for PCR using E. coli atoS gene-specific primer pairs. Lanes: 1, without dilution; 2, 10 −1 dilution; 3, 10 −2 dilution; 4, 10 −3 dilution; 5, 10 −4 dilution; 6, 10 −5 dilution; 7, 10 −6 dilution.
    Biofilms, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher streptavidin coated 96 well maxisorp plates
    ), are shown. (B) Peptide A specificity of the MAbs in an ELISA. Biotin-conjugated peptide A was added to <t>streptavidin-coated</t> 96-well plates (200 ng/well). Each MAb (ascites fluid) was diluted 1:1,000 and used as the primary antibody. The y . An ELISA was performed with 1:20,000-diluted ascites fluid in the presence or absence of various concentrations of NaSCN as indicated. The specific binding affinity was calculated based on the values obtained with or without NaSCN. The data shown represent three independent experiments. Error bars represent the standard deviation.
    Streptavidin Coated 96 Well Maxisorp Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher viral genomic rna
    ABT-263 induces the premature death of cells <t>transfected</t> with IAV genomic <t>RNA</t> (vRNA) or plasmid DNA (pDNA). ( A ) Fluorescent microscopy images showing that ABT-263 kills vRNA-transfected (160 ng) but not mock-transfected RPE cells at 8 h post transfection. Asymmetric cyanine dye stains the dsDNA of dead cells. Hoechst stains DNA in living cells; ( B ) CTxG plot showing that ABT-263 (3 µM) induces that premature death of RPE cells transfected with increasing concentrations of vRNA. Mean ± SD, n = 3; ( C ) Fluorescent and bright field microscopy of RPE cells showing that ABT-263 kills eGFP-expressing plasmid transfected (300 ng) but not mock-transfected RPE cells at 6 h post transfection; ( D ) CTG graph showing that the viability of ABT-263-treated (3 µM) cells decreases with increasing concentrations of transfected plasmid DNA. Mean ± SD, n = 3.
    Viral Genomic Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher 96 well pcr plates
    Workflow outlining the major steps of the study. SYBR Green I-stained single sperm cells were isolated by fluorescence-activated cell sorting (FACS) to <t>96-well</t> <t>PCR</t> plates; DNA was amplified and used for next-generation sequencing
    96 Well Pcr Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Loss of outer membrane integrity in strain RN102. Bacterial samples were collected from 48-hour-cultured biofilms for TEM analysis. TEM images of the bacterial cells and the cell appendages are shown for strains: (A) BW25113, (B) RN102, (C) BW25113/pNTR-SD, (D) RN102/pNTR-SD, and (E) RN102/pNT3( hldE ). The outer membranes are indicated by arrows. Representative electron-microphotographs of each strain are shown. A 500-nm-long bar is shown in the lower left corner of each eclectron-micrograph. (F) Western blot analysis of supernatants from BW25113 and RN102. Supernatants were harvested by centrifugation from bacterial liquid culture grown for 48 hours under static conditions. Results of Western blot using anti-Crp, anti-DsbA, anti-OmpC, and anti-OmpA antisera are shown. (G) Supernatants from bacterial liquid cultures of BW25113 or RN102 grown for 48 hours under static conditions were serially diluted with TE. The diluted samples were used as template DNA for PCR using E. coli atoS gene-specific primer pairs. Lanes: 1, without dilution; 2, 10 −1 dilution; 3, 10 −2 dilution; 4, 10 −3 dilution; 5, 10 −4 dilution; 6, 10 −5 dilution; 7, 10 −6 dilution.

    Journal: PLoS ONE

    Article Title: Enhanced Biofilm Formation by Escherichia coli LPS Mutants Defective in Hep Biosynthesis

    doi: 10.1371/journal.pone.0051241

    Figure Lengend Snippet: Loss of outer membrane integrity in strain RN102. Bacterial samples were collected from 48-hour-cultured biofilms for TEM analysis. TEM images of the bacterial cells and the cell appendages are shown for strains: (A) BW25113, (B) RN102, (C) BW25113/pNTR-SD, (D) RN102/pNTR-SD, and (E) RN102/pNT3( hldE ). The outer membranes are indicated by arrows. Representative electron-microphotographs of each strain are shown. A 500-nm-long bar is shown in the lower left corner of each eclectron-micrograph. (F) Western blot analysis of supernatants from BW25113 and RN102. Supernatants were harvested by centrifugation from bacterial liquid culture grown for 48 hours under static conditions. Results of Western blot using anti-Crp, anti-DsbA, anti-OmpC, and anti-OmpA antisera are shown. (G) Supernatants from bacterial liquid cultures of BW25113 or RN102 grown for 48 hours under static conditions were serially diluted with TE. The diluted samples were used as template DNA for PCR using E. coli atoS gene-specific primer pairs. Lanes: 1, without dilution; 2, 10 −1 dilution; 3, 10 −2 dilution; 4, 10 −3 dilution; 5, 10 −4 dilution; 6, 10 −5 dilution; 7, 10 −6 dilution.

    Article Snippet: In order to quantify the amount of biofilm on a 96-well plate, all stain associated with the attached biofilms was dissolved with 95% ethanol, then OD595 absorbance was measured using a microplate reader (Multiskan RC, ThermoFisher, Waltham, MA).

    Techniques: Cell Culture, Transmission Electron Microscopy, Western Blot, Centrifugation, Polymerase Chain Reaction

    Contribution of eDNA to biofilm structure formed by RN102. (A–E) CLSM images of biofilms formed by strains: (A) BW25113 (B) RN102, (C) BW25113/pNTR-SD, (D) RN102/pNTR-SD, and (D) RN102/pNT3( hldE ). Images of biofilms stained with acrydine orange are shown as digital CLSM images. In each strain, a section which has the largest sum of signals in the defined area (127.3 μm by 127.3 μm) among all X–Y sections is shown in the upper row (X–Y). The overview of biofilms in the same area of each X–Y section is shown as 3D image in the lower row (3D). The volume of each 3D image (μm 3 ) in the area of the X–Y planes was quantified and the mean ± SD obtained from 3 different areas chosen at random are denoted in the upper-right corners. The data shown are representative microphotographs of two independent experiments. (F) Quantification of eDNA from BW25113 and RN102 strains. The bars represent the ratio of extracellular DNA to intracellular DNA (eDNA/iDNA). Results are shown as the mean ± SD from 3 independent experiments. * P

    Journal: PLoS ONE

    Article Title: Enhanced Biofilm Formation by Escherichia coli LPS Mutants Defective in Hep Biosynthesis

    doi: 10.1371/journal.pone.0051241

    Figure Lengend Snippet: Contribution of eDNA to biofilm structure formed by RN102. (A–E) CLSM images of biofilms formed by strains: (A) BW25113 (B) RN102, (C) BW25113/pNTR-SD, (D) RN102/pNTR-SD, and (D) RN102/pNT3( hldE ). Images of biofilms stained with acrydine orange are shown as digital CLSM images. In each strain, a section which has the largest sum of signals in the defined area (127.3 μm by 127.3 μm) among all X–Y sections is shown in the upper row (X–Y). The overview of biofilms in the same area of each X–Y section is shown as 3D image in the lower row (3D). The volume of each 3D image (μm 3 ) in the area of the X–Y planes was quantified and the mean ± SD obtained from 3 different areas chosen at random are denoted in the upper-right corners. The data shown are representative microphotographs of two independent experiments. (F) Quantification of eDNA from BW25113 and RN102 strains. The bars represent the ratio of extracellular DNA to intracellular DNA (eDNA/iDNA). Results are shown as the mean ± SD from 3 independent experiments. * P

    Article Snippet: In order to quantify the amount of biofilm on a 96-well plate, all stain associated with the attached biofilms was dissolved with 95% ethanol, then OD595 absorbance was measured using a microplate reader (Multiskan RC, ThermoFisher, Waltham, MA).

    Techniques: Confocal Laser Scanning Microscopy, Staining

    Biofilm formation and growth of a series of LPS mutants. (A) Biofilm formation by a series of core OS LPS mutants when compared to the parental strain, BW25113. The mean ± SD of results from 3 independent experiments are shown. Statistical analysis was performed using ANOVA. * P

    Journal: PLoS ONE

    Article Title: Enhanced Biofilm Formation by Escherichia coli LPS Mutants Defective in Hep Biosynthesis

    doi: 10.1371/journal.pone.0051241

    Figure Lengend Snippet: Biofilm formation and growth of a series of LPS mutants. (A) Biofilm formation by a series of core OS LPS mutants when compared to the parental strain, BW25113. The mean ± SD of results from 3 independent experiments are shown. Statistical analysis was performed using ANOVA. * P

    Article Snippet: In order to quantify the amount of biofilm on a 96-well plate, all stain associated with the attached biofilms was dissolved with 95% ethanol, then OD595 absorbance was measured using a microplate reader (Multiskan RC, ThermoFisher, Waltham, MA).

    Techniques:

    Loss of flagella in RN102. Fourty eight-hour-cultured biofilms were collected and analyzed by TEM, and by Western blot for FliC. (A–E) TEM images of the bacterial cells and the cell appendages are shown for strains (A) BW25113, (B) RN102, (C) BW25113/pNTR-SD, (D) RN102/pNTR-SD, and (E) RN102/pNT3( hldE ). Flagella found in figures (A), (C), and (E) are shown by arrowheads. Representative electron-microphotographs of each strain are shown. A 1-μm-long bar is shown in the lower left corner. (F) Supernatants were collected from 48-hour bacterial cultures. Result of Western blot using anti-FliC antiserum is shown. Lanes; 1, BW25113; 2, RN102; 3, BW25113/pNTR-SD; 4, RN102/pNTR-SD; 5, RN102/pNT3( hldE ); 6, RN110.

    Journal: PLoS ONE

    Article Title: Enhanced Biofilm Formation by Escherichia coli LPS Mutants Defective in Hep Biosynthesis

    doi: 10.1371/journal.pone.0051241

    Figure Lengend Snippet: Loss of flagella in RN102. Fourty eight-hour-cultured biofilms were collected and analyzed by TEM, and by Western blot for FliC. (A–E) TEM images of the bacterial cells and the cell appendages are shown for strains (A) BW25113, (B) RN102, (C) BW25113/pNTR-SD, (D) RN102/pNTR-SD, and (E) RN102/pNT3( hldE ). Flagella found in figures (A), (C), and (E) are shown by arrowheads. Representative electron-microphotographs of each strain are shown. A 1-μm-long bar is shown in the lower left corner. (F) Supernatants were collected from 48-hour bacterial cultures. Result of Western blot using anti-FliC antiserum is shown. Lanes; 1, BW25113; 2, RN102; 3, BW25113/pNTR-SD; 4, RN102/pNTR-SD; 5, RN102/pNT3( hldE ); 6, RN110.

    Article Snippet: In order to quantify the amount of biofilm on a 96-well plate, all stain associated with the attached biofilms was dissolved with 95% ethanol, then OD595 absorbance was measured using a microplate reader (Multiskan RC, ThermoFisher, Waltham, MA).

    Techniques: Cell Culture, Transmission Electron Microscopy, Western Blot

    ), are shown. (B) Peptide A specificity of the MAbs in an ELISA. Biotin-conjugated peptide A was added to streptavidin-coated 96-well plates (200 ng/well). Each MAb (ascites fluid) was diluted 1:1,000 and used as the primary antibody. The y . An ELISA was performed with 1:20,000-diluted ascites fluid in the presence or absence of various concentrations of NaSCN as indicated. The specific binding affinity was calculated based on the values obtained with or without NaSCN. The data shown represent three independent experiments. Error bars represent the standard deviation.

    Journal: Journal of Virology

    Article Title: Amino Acid Residue-Specific Neutralization and Nonneutralization of Hepatitis C Virus by Monoclonal Antibodies to the E2 Protein

    doi: 10.1128/JVI.00994-12

    Figure Lengend Snippet: ), are shown. (B) Peptide A specificity of the MAbs in an ELISA. Biotin-conjugated peptide A was added to streptavidin-coated 96-well plates (200 ng/well). Each MAb (ascites fluid) was diluted 1:1,000 and used as the primary antibody. The y . An ELISA was performed with 1:20,000-diluted ascites fluid in the presence or absence of various concentrations of NaSCN as indicated. The specific binding affinity was calculated based on the values obtained with or without NaSCN. The data shown represent three independent experiments. Error bars represent the standard deviation.

    Article Snippet: Biotin-conjugated peptide (200 ng/well) was added to streptavidin-coated 96-well Maxisorp plates (Thermo Fisher Scientific, Rockford, IL), followed by incubation at room temperature for 1 h in Super Block blocking buffer (Thermo Scientific).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Standard Deviation

    Use of mutational analysis to identify the residues that are critical for antibody recognition. (A) Biotin-conjugated peptides were chemically synthesized to represent the truncated peptide B, i.e., B short, from the E2 protein of HCV genotype 1a (H77 strain) and its mutations. The B short mutant peptides contained a single alanine substitution at positions 437, 438, 440, 441, and 442, respectively. A hyphen indicates an amino acid residue identical to that of the H77 sequence. (B) Biotin-conjugated B short peptide and its mutants were added to streptavidin-coated 96-well plates at 200 ng/well in an ELISA. Each MAb (ascites fluid) was diluted 1:10 5 dilution, and applied as the primary antibody. The data shown represent at least three independent experiments. The x axis indicates the antibodies used in the assay. The y axis indicates the absorbance at 405 nm, representing specific binding of a given antibody to each individual peptide.

    Journal: Journal of Virology

    Article Title: Amino Acid Residue-Specific Neutralization and Nonneutralization of Hepatitis C Virus by Monoclonal Antibodies to the E2 Protein

    doi: 10.1128/JVI.00994-12

    Figure Lengend Snippet: Use of mutational analysis to identify the residues that are critical for antibody recognition. (A) Biotin-conjugated peptides were chemically synthesized to represent the truncated peptide B, i.e., B short, from the E2 protein of HCV genotype 1a (H77 strain) and its mutations. The B short mutant peptides contained a single alanine substitution at positions 437, 438, 440, 441, and 442, respectively. A hyphen indicates an amino acid residue identical to that of the H77 sequence. (B) Biotin-conjugated B short peptide and its mutants were added to streptavidin-coated 96-well plates at 200 ng/well in an ELISA. Each MAb (ascites fluid) was diluted 1:10 5 dilution, and applied as the primary antibody. The data shown represent at least three independent experiments. The x axis indicates the antibodies used in the assay. The y axis indicates the absorbance at 405 nm, representing specific binding of a given antibody to each individual peptide.

    Article Snippet: Biotin-conjugated peptide (200 ng/well) was added to streptavidin-coated 96-well Maxisorp plates (Thermo Fisher Scientific, Rockford, IL), followed by incubation at room temperature for 1 h in Super Block blocking buffer (Thermo Scientific).

    Techniques: Synthesized, Mutagenesis, Sequencing, Enzyme-linked Immunosorbent Assay, Binding Assay

    ABT-263 induces the premature death of cells transfected with IAV genomic RNA (vRNA) or plasmid DNA (pDNA). ( A ) Fluorescent microscopy images showing that ABT-263 kills vRNA-transfected (160 ng) but not mock-transfected RPE cells at 8 h post transfection. Asymmetric cyanine dye stains the dsDNA of dead cells. Hoechst stains DNA in living cells; ( B ) CTxG plot showing that ABT-263 (3 µM) induces that premature death of RPE cells transfected with increasing concentrations of vRNA. Mean ± SD, n = 3; ( C ) Fluorescent and bright field microscopy of RPE cells showing that ABT-263 kills eGFP-expressing plasmid transfected (300 ng) but not mock-transfected RPE cells at 6 h post transfection; ( D ) CTG graph showing that the viability of ABT-263-treated (3 µM) cells decreases with increasing concentrations of transfected plasmid DNA. Mean ± SD, n = 3.

    Journal: Viruses

    Article Title: Antiviral Properties of Chemical Inhibitors of Cellular Anti-Apoptotic Bcl-2 Proteins

    doi: 10.3390/v9100271

    Figure Lengend Snippet: ABT-263 induces the premature death of cells transfected with IAV genomic RNA (vRNA) or plasmid DNA (pDNA). ( A ) Fluorescent microscopy images showing that ABT-263 kills vRNA-transfected (160 ng) but not mock-transfected RPE cells at 8 h post transfection. Asymmetric cyanine dye stains the dsDNA of dead cells. Hoechst stains DNA in living cells; ( B ) CTxG plot showing that ABT-263 (3 µM) induces that premature death of RPE cells transfected with increasing concentrations of vRNA. Mean ± SD, n = 3; ( C ) Fluorescent and bright field microscopy of RPE cells showing that ABT-263 kills eGFP-expressing plasmid transfected (300 ng) but not mock-transfected RPE cells at 6 h post transfection; ( D ) CTG graph showing that the viability of ABT-263-treated (3 µM) cells decreases with increasing concentrations of transfected plasmid DNA. Mean ± SD, n = 3.

    Article Snippet: Transfections of RPE Cells with vRNA or Plasmid DNA RPE cells were cultured to 80% confluence in 96 well plates and transfected with 160 ng viral genomic RNA using Lipofectamine RNAiMAX (Thermo Fisher Scientific, Waltham, MA, USA) or with 30, 100, or 300 ng of plasmid DNA (pEGFP) using Lipofectamine 3000 (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Transfection, Plasmid Preparation, Microscopy, Expressing, CTG Assay

    Workflow outlining the major steps of the study. SYBR Green I-stained single sperm cells were isolated by fluorescence-activated cell sorting (FACS) to 96-well PCR plates; DNA was amplified and used for next-generation sequencing

    Journal: Journal of Assisted Reproduction and Genetics

    Article Title: Chromosomal scan of single sperm cells by combining fluorescence-activated cell sorting and next-generation sequencing

    doi: 10.1007/s10815-018-1340-0

    Figure Lengend Snippet: Workflow outlining the major steps of the study. SYBR Green I-stained single sperm cells were isolated by fluorescence-activated cell sorting (FACS) to 96-well PCR plates; DNA was amplified and used for next-generation sequencing

    Article Snippet: Single sperms were collected into 96-well PCR plates (Thermo Fisher Scientific, USA) containing 3 μL PBS per well.

    Techniques: SYBR Green Assay, Staining, Isolation, Fluorescence, FACS, Polymerase Chain Reaction, Amplification, Next-Generation Sequencing