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  • 99
    Thermo Fisher biolite multidishes and microwell plates
    Biolite Multidishes And Microwell Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 12 well plates
    Conversion of LC3B-I to LC3B-II and morphology of autophagosomes in HeLa cells with KD of FHF subunits. HeLa cells were subjected to two rounds of treatment with the corresponding siRNA pools as described in Materials and Methods . (A) IB analysis of LC3B-I and LC3B-II in control and siRNA-treated cells seeded on <t>12-well</t> plates and subjected to amino acid and serum starvation for 45 min at 37°C (image at left) or treatment with 100 nM bafilomycin A1 for 4 h at 37°C (image at right). IB for α-tubulin is shown as loading control. (B–D) Cells seeded on glass coverslips were fixed with methanol at –20°C and immunostained for endogenous LC3B, AP-4 ε, and TGN46 followed by confocal microscopy imaging. Anti-LC3B, anti-AP-4 ε, and anti-TGN46 immunostaining is shown in green, red, and blue, respectively. The panels at right depict merge images with DAPI staining of nuclei in gray (cells are outlined by dashed white lines). Arrows in C and D (left column) point at tubular or “thread-like” autophagosomes or large autophagosomes detected in a fraction of FHIP KD and FHIP-L KD cells, respectively (an enlargement of the area inside the white box in C is shown at the top right of the merge image). Scale bars: 5 μm for the enlargement in C (right column) and 10 μm for all other images. Cells were manually scored for the presence of LC3B-positive structures on threads or of large LC3B-positive structures (approximately 0.9–1.6 μm diameter range). We observed that 4.3 ± 0.7% of FHIP KD cells exhibited autophagosomes on threads compared with 0.3 ± 0.2% in control cells and 0 ± 0% in FHIP-L KD cells (mean ± SEM, P
    12 Well Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 597 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Corning Life Sciences 12 well plates
    Detection of the viral protein in supernatants and lysates of IAV-infected cells. MDCK cells plated in <t>12-well</t> plates were infected with IAVs at m.o.i. 2.0 and incubated with or without MAbs B12 and ch-rM2ss23 for 8 h at 35 °C. The cells infected with Adachi and Aichi were incubated with or without the MAbs at 10 µg/mL and 1 µg/mL, respectively. The M1 protein in supernatants ( a ) and cell lysates ( b ) was detected in western blotting. Beta-actin was also stained for cell lysate samples. Band intensities relative to each control sample (i.e., cells incubated without MAbs) are shown. Each experiment was performed three times and averages and standard deviations are shown. Asterisks indicate significant differences (* p
    12 Well Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 528 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Corning Life Sciences 12 well tissue culture plates
    Effect of SMase-LDL and cysteamine (Cys) on LysoTracker Red accumulation by macrophages. THP-1 macrophages or HMDMs (1 × 10 6 ) were cultured in <t>12-well</t> tissue culture plates in RPMI medium (containing 10% v/v FCS) alone or containing native LDL or SMase-LDL with or without cysteamine (10 or 25 μM) for 72 h. All LDL concentrations were 100 μg protein/ml. After 72 h, cells were treated with 500 nM LysoTracker Red for 30 min and then assayed by flow cytometry. The MFI peak of LysoTracker Red in the red channel was then measured. A: MFI in red channel of healthy THP-1 macrophages, native-LDL-treated macrophages, and SMase-LDL-treated macrophages. B: MFI in red channel of SMase-LDL treated THP-1 macrophages in the presence or absence of cysteamine (10 and 25 μM). C: Data expressed as percentage loss of MFI of LysoTracker Red in the red channel compared with untreated control macrophages in THP-1 macrophages. D: Data expressed as percentage loss of MFI of LysoTracker Red in the red channel compared with untreated macrophages in HMDMs. ** P
    12 Well Tissue Culture Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 419 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson 12 well tissue culture plates
    Effect of 1,25(OH) 2 D3 on estrogen-induced proliferation of cultured human UF cells. A, HuLM cells (5000 cells/well) in <t>12-well</t> plates were cultured in phenol free smooth muscle basal medium containing 5% fetal bovine serum. Cells were treated with 10
    12 Well Tissue Culture Plates, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 358 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher nunc 12 well carrier plate for cell culture inserts
    Effect of 1,25(OH) 2 D3 on estrogen-induced proliferation of cultured human UF cells. A, HuLM cells (5000 cells/well) in <t>12-well</t> plates were cultured in phenol free smooth muscle basal medium containing 5% fetal bovine serum. Cells were treated with 10
    Nunc 12 Well Carrier Plate For Cell Culture Inserts, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Corning Life Sciences 12 well transwell plates
    Evaluation of the BBB-penetration ability of CPDs in vitro. (A–C) Schematic diagram of the preparation and application of the in vitro BBB model: (A) the <t>12-well</t> plates were seeded with C6 cells; (B) HUVEC were seeded into the <t>transwell</t> inserts; (C) CPDs were added into the transwell lumen, and the fluid in the lower compartment containing the CPDs could be detected. (D) Time-dependent TEER values of the in vitro BBB model ( n = 3, mean ± SD). (E) Accumulated percentages of CPDs that crossed the BBB ( n = 6, mean ± SD).
    12 Well Transwell Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 89/100, based on 192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson 12 well culture plates
    Evaluation of the BBB-penetration ability of CPDs in vitro. (A–C) Schematic diagram of the preparation and application of the in vitro BBB model: (A) the <t>12-well</t> plates were seeded with C6 cells; (B) HUVEC were seeded into the <t>transwell</t> inserts; (C) CPDs were added into the transwell lumen, and the fluid in the lower compartment containing the CPDs could be detected. (D) Time-dependent TEER values of the in vitro BBB model ( n = 3, mean ± SD). (E) Accumulated percentages of CPDs that crossed the BBB ( n = 6, mean ± SD).
    12 Well Culture Plates, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 12 well culture plates
    Formation of syncytia by CC81 cells transfected with infectious BLV clones. CC81 cells were transfected with infectious clones of BLV or the control plasmid. Then, 24 h after transfection, transfected cells were replated at 8 × 10 4 cells/well in <t>12-well</t> culture plates and cultured for 2 days in complete medium with 4 mg of Polybrene per ml. The cells were fixed and stained, and the syncytia and nuclei in each syncytium were counted. Syncytia were classified into three groups: syncytia with 5 to 9 nuclei (left), 10 to 14 nuclei (center), and 15 or more nuclei (right). Average values from three wells with standard error (error bars) are shown.
    12 Well Culture Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 236 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Greiner Bio 12 well plates
    Drawing Voronoi diagrams, and examples of their use. A. Principle. ( i ) In this example (4 points randomly-distributed in a square), ( ii ) we draw straight lines between each point (here shown for point 2) and its nearest neighbors. ( iii ) The corresponding Voronoi polygon is formed using half-plane intersections for all lines. ( iv ) The same procedure is applied for other points to produce the Voronoi diagram. B, C. Example Voronoi diagrams used during cloning of ES and HEK cells. Details are as Fig. 3 , and each pair of images on the left show the same region of the dish immediately after plating cells (d 0; circles surround single cells), and prior to picking colonies (d 7; dotted lines outline colonies). Right-hand panels show views of picked colonies growing in conventional <t>12-well</t> plates at d 17.
    12 Well Plates, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 93/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 12 well culture plates
    Drawing Voronoi diagrams, and examples of their use. A. Principle. ( i ) In this example (4 points randomly-distributed in a square), ( ii ) we draw straight lines between each point (here shown for point 2) and its nearest neighbors. ( iii ) The corresponding Voronoi polygon is formed using half-plane intersections for all lines. ( iv ) The same procedure is applied for other points to produce the Voronoi diagram. B, C. Example Voronoi diagrams used during cloning of ES and HEK cells. Details are as Fig. 3 , and each pair of images on the left show the same region of the dish immediately after plating cells (d 0; circles surround single cells), and prior to picking colonies (d 7; dotted lines outline colonies). Right-hand panels show views of picked colonies growing in conventional <t>12-well</t> plates at d 17.
    12 Well Culture Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 473 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Corning Life Sciences 12 well transwell inserts
    Co-culture model description. IEC were grown in <t>12-well</t> transwell inserts until confluency and basolaterally exposed to either non-activated or αCD3/CD28-activated PBMC. Apically, IEC were conditioned with 2′-FL or GF in the presence or absence of CpG, a TLR9 agonist mimicking a bacterial trigger ( A ). After 24 h incubation, basolateral supernatant was collected to analyze the T-cell mediator release. The IEC were set apart and washed with PBS. Then, fresh medium was added and IEC were kept in incubation for an additional 24 h to study the IEC-derived mediator release ( B ). Alternatively, IEC were washed with PBS and co-cultured with immature moDC for 48 h ( C ). Then, the basolateral supernatant was collected where the mediator release was studied. Additionally, the phenotype of moDC after IEC/moDC co-culture was analyzed. Subsequently, conditioned moDC (ccDC) were exposed to naïve T-cells in an allogeneic DC/T-cell assay ( D ). After 5–6 days incubation, the cytokine release was measured in the supernatant.
    12 Well Transwell Inserts, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 91/100, based on 118 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    MatTek glass bottom 12 well plates
    Co-culture model description. IEC were grown in <t>12-well</t> transwell inserts until confluency and basolaterally exposed to either non-activated or αCD3/CD28-activated PBMC. Apically, IEC were conditioned with 2′-FL or GF in the presence or absence of CpG, a TLR9 agonist mimicking a bacterial trigger ( A ). After 24 h incubation, basolateral supernatant was collected to analyze the T-cell mediator release. The IEC were set apart and washed with PBS. Then, fresh medium was added and IEC were kept in incubation for an additional 24 h to study the IEC-derived mediator release ( B ). Alternatively, IEC were washed with PBS and co-cultured with immature moDC for 48 h ( C ). Then, the basolateral supernatant was collected where the mediator release was studied. Additionally, the phenotype of moDC after IEC/moDC co-culture was analyzed. Subsequently, conditioned moDC (ccDC) were exposed to naïve T-cells in an allogeneic DC/T-cell assay ( D ). After 5–6 days incubation, the cytokine release was measured in the supernatant.
    Glass Bottom 12 Well Plates, supplied by MatTek, used in various techniques. Bioz Stars score: 88/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 12 well tissue culture plates
    Co-culture model description. IEC were grown in <t>12-well</t> transwell inserts until confluency and basolaterally exposed to either non-activated or αCD3/CD28-activated PBMC. Apically, IEC were conditioned with 2′-FL or GF in the presence or absence of CpG, a TLR9 agonist mimicking a bacterial trigger ( A ). After 24 h incubation, basolateral supernatant was collected to analyze the T-cell mediator release. The IEC were set apart and washed with PBS. Then, fresh medium was added and IEC were kept in incubation for an additional 24 h to study the IEC-derived mediator release ( B ). Alternatively, IEC were washed with PBS and co-cultured with immature moDC for 48 h ( C ). Then, the basolateral supernatant was collected where the mediator release was studied. Additionally, the phenotype of moDC after IEC/moDC co-culture was analyzed. Subsequently, conditioned moDC (ccDC) were exposed to naïve T-cells in an allogeneic DC/T-cell assay ( D ). After 5–6 days incubation, the cytokine release was measured in the supernatant.
    12 Well Tissue Culture Plates, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 241 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Conversion of LC3B-I to LC3B-II and morphology of autophagosomes in HeLa cells with KD of FHF subunits. HeLa cells were subjected to two rounds of treatment with the corresponding siRNA pools as described in Materials and Methods . (A) IB analysis of LC3B-I and LC3B-II in control and siRNA-treated cells seeded on 12-well plates and subjected to amino acid and serum starvation for 45 min at 37°C (image at left) or treatment with 100 nM bafilomycin A1 for 4 h at 37°C (image at right). IB for α-tubulin is shown as loading control. (B–D) Cells seeded on glass coverslips were fixed with methanol at –20°C and immunostained for endogenous LC3B, AP-4 ε, and TGN46 followed by confocal microscopy imaging. Anti-LC3B, anti-AP-4 ε, and anti-TGN46 immunostaining is shown in green, red, and blue, respectively. The panels at right depict merge images with DAPI staining of nuclei in gray (cells are outlined by dashed white lines). Arrows in C and D (left column) point at tubular or “thread-like” autophagosomes or large autophagosomes detected in a fraction of FHIP KD and FHIP-L KD cells, respectively (an enlargement of the area inside the white box in C is shown at the top right of the merge image). Scale bars: 5 μm for the enlargement in C (right column) and 10 μm for all other images. Cells were manually scored for the presence of LC3B-positive structures on threads or of large LC3B-positive structures (approximately 0.9–1.6 μm diameter range). We observed that 4.3 ± 0.7% of FHIP KD cells exhibited autophagosomes on threads compared with 0.3 ± 0.2% in control cells and 0 ± 0% in FHIP-L KD cells (mean ± SEM, P

    Journal: Molecular Biology of the Cell

    Article Title: The FTS-Hook-FHIP (FHF) complex interacts with AP-4 to mediate perinuclear distribution of AP-4 and its cargo ATG9A

    doi: 10.1091/mbc.E19-11-0658

    Figure Lengend Snippet: Conversion of LC3B-I to LC3B-II and morphology of autophagosomes in HeLa cells with KD of FHF subunits. HeLa cells were subjected to two rounds of treatment with the corresponding siRNA pools as described in Materials and Methods . (A) IB analysis of LC3B-I and LC3B-II in control and siRNA-treated cells seeded on 12-well plates and subjected to amino acid and serum starvation for 45 min at 37°C (image at left) or treatment with 100 nM bafilomycin A1 for 4 h at 37°C (image at right). IB for α-tubulin is shown as loading control. (B–D) Cells seeded on glass coverslips were fixed with methanol at –20°C and immunostained for endogenous LC3B, AP-4 ε, and TGN46 followed by confocal microscopy imaging. Anti-LC3B, anti-AP-4 ε, and anti-TGN46 immunostaining is shown in green, red, and blue, respectively. The panels at right depict merge images with DAPI staining of nuclei in gray (cells are outlined by dashed white lines). Arrows in C and D (left column) point at tubular or “thread-like” autophagosomes or large autophagosomes detected in a fraction of FHIP KD and FHIP-L KD cells, respectively (an enlargement of the area inside the white box in C is shown at the top right of the merge image). Scale bars: 5 μm for the enlargement in C (right column) and 10 μm for all other images. Cells were manually scored for the presence of LC3B-positive structures on threads or of large LC3B-positive structures (approximately 0.9–1.6 μm diameter range). We observed that 4.3 ± 0.7% of FHIP KD cells exhibited autophagosomes on threads compared with 0.3 ± 0.2% in control cells and 0 ± 0% in FHIP-L KD cells (mean ± SEM, P

    Article Snippet: Amino acid, serum starvation, and bafilomycin treatment Control and siRNA-treated cells were seeded on 12-well plates the day before the experiment.

    Techniques: Confocal Microscopy, Imaging, Immunostaining, Staining

    Detection of the viral protein in supernatants and lysates of IAV-infected cells. MDCK cells plated in 12-well plates were infected with IAVs at m.o.i. 2.0 and incubated with or without MAbs B12 and ch-rM2ss23 for 8 h at 35 °C. The cells infected with Adachi and Aichi were incubated with or without the MAbs at 10 µg/mL and 1 µg/mL, respectively. The M1 protein in supernatants ( a ) and cell lysates ( b ) was detected in western blotting. Beta-actin was also stained for cell lysate samples. Band intensities relative to each control sample (i.e., cells incubated without MAbs) are shown. Each experiment was performed three times and averages and standard deviations are shown. Asterisks indicate significant differences (* p

    Journal: Viruses

    Article Title: Comparative Analyses of the Antiviral Activities of IgG and IgA Antibodies to Influenza A Virus M2 Protein

    doi: 10.3390/v12070780

    Figure Lengend Snippet: Detection of the viral protein in supernatants and lysates of IAV-infected cells. MDCK cells plated in 12-well plates were infected with IAVs at m.o.i. 2.0 and incubated with or without MAbs B12 and ch-rM2ss23 for 8 h at 35 °C. The cells infected with Adachi and Aichi were incubated with or without the MAbs at 10 µg/mL and 1 µg/mL, respectively. The M1 protein in supernatants ( a ) and cell lysates ( b ) was detected in western blotting. Beta-actin was also stained for cell lysate samples. Band intensities relative to each control sample (i.e., cells incubated without MAbs) are shown. Each experiment was performed three times and averages and standard deviations are shown. Asterisks indicate significant differences (* p

    Article Snippet: Sample Collection of Cell Lysates and Supernatants of Infected CellsMDCK cells seeded on 12-well plates (Corning, Corning, NY, USA) were incubated with IAVs at a multiplicity of infection (m.o.i.) of 2.0 for one hour, and then washed with PBS.

    Techniques: Infection, Incubation, Western Blot, Staining

    Growth analysis of mESC lines. All mESCs were maintained and seeded with equal cell concentrations on either a MEF feeder layers (Passage 3–5) or gelatin (Passage 6–8). ( A)  Days to confluence. The longer time to confluence in all cell lines on passage 5 (Fig. 1A) was secondary to use larger well size starting from passage 4 (cells were moved from a 24 well plate with a diameter of 15 mm on passage 1 to 3, to a 12 well plate with a diameter of 20 mm on passage 4 and 5 to a well plate with a diameter of 35 mm in passages 6–8). ( B ) Percentage unattached cells after 12h plating. ( C ) Doubling time. Arrow indicates passage to 12 well plates. Arrowhead indicates passage on gelatin.

    Journal: PLoS ONE

    Article Title: Embryonic Stem Cells Derived from In Vivo or In Vitro-Generated Murine Blastocysts Display Similar Transcriptome and Differentiation Potential

    doi: 10.1371/journal.pone.0117422

    Figure Lengend Snippet: Growth analysis of mESC lines. All mESCs were maintained and seeded with equal cell concentrations on either a MEF feeder layers (Passage 3–5) or gelatin (Passage 6–8). ( A) Days to confluence. The longer time to confluence in all cell lines on passage 5 (Fig. 1A) was secondary to use larger well size starting from passage 4 (cells were moved from a 24 well plate with a diameter of 15 mm on passage 1 to 3, to a 12 well plate with a diameter of 20 mm on passage 4 and 5 to a well plate with a diameter of 35 mm in passages 6–8). ( B ) Percentage unattached cells after 12h plating. ( C ) Doubling time. Arrow indicates passage to 12 well plates. Arrowhead indicates passage on gelatin.

    Article Snippet: After trypsinization, equal cell concentrations were plated onto 12-well culture plates (Corning) and the number of days to reach confluence, the doubling time and percent cell attachment were monitored.

    Techniques:

    Effect of SMase-LDL and cysteamine (Cys) on LysoTracker Red accumulation by macrophages. THP-1 macrophages or HMDMs (1 × 10 6 ) were cultured in 12-well tissue culture plates in RPMI medium (containing 10% v/v FCS) alone or containing native LDL or SMase-LDL with or without cysteamine (10 or 25 μM) for 72 h. All LDL concentrations were 100 μg protein/ml. After 72 h, cells were treated with 500 nM LysoTracker Red for 30 min and then assayed by flow cytometry. The MFI peak of LysoTracker Red in the red channel was then measured. A: MFI in red channel of healthy THP-1 macrophages, native-LDL-treated macrophages, and SMase-LDL-treated macrophages. B: MFI in red channel of SMase-LDL treated THP-1 macrophages in the presence or absence of cysteamine (10 and 25 μM). C: Data expressed as percentage loss of MFI of LysoTracker Red in the red channel compared with untreated control macrophages in THP-1 macrophages. D: Data expressed as percentage loss of MFI of LysoTracker Red in the red channel compared with untreated macrophages in HMDMs. ** P

    Journal: Journal of Lipid Research

    Article Title: Lysosomal oxidation of LDL alters lysosomal pH, induces senescence, and increases secretion of pro-inflammatory cytokines in human macrophages [S]

    doi: 10.1194/jlr.M088245

    Figure Lengend Snippet: Effect of SMase-LDL and cysteamine (Cys) on LysoTracker Red accumulation by macrophages. THP-1 macrophages or HMDMs (1 × 10 6 ) were cultured in 12-well tissue culture plates in RPMI medium (containing 10% v/v FCS) alone or containing native LDL or SMase-LDL with or without cysteamine (10 or 25 μM) for 72 h. All LDL concentrations were 100 μg protein/ml. After 72 h, cells were treated with 500 nM LysoTracker Red for 30 min and then assayed by flow cytometry. The MFI peak of LysoTracker Red in the red channel was then measured. A: MFI in red channel of healthy THP-1 macrophages, native-LDL-treated macrophages, and SMase-LDL-treated macrophages. B: MFI in red channel of SMase-LDL treated THP-1 macrophages in the presence or absence of cysteamine (10 and 25 μM). C: Data expressed as percentage loss of MFI of LysoTracker Red in the red channel compared with untreated control macrophages in THP-1 macrophages. D: Data expressed as percentage loss of MFI of LysoTracker Red in the red channel compared with untreated macrophages in HMDMs. ** P

    Article Snippet: THP-1 macrophages or HMDMs (4,000 per well) were plated in 12-well tissue culture plates (Corning).

    Techniques: Cell Culture, Flow Cytometry, Cytometry

    Effect of lysosomal oxidation of LDL on senescence in human macrophages. HMDMs were cultured in 12-well tissue culture plates at 3,000 cells per well in RPMI medium (containing 10% v/v lipoprotein-deficient serum) containing either no LDL (A), native LDL (B), SMase-LDL alone (C), or SMase-LDL (D) (all at 100 μg protein/ml) with 10 μM cysteamine (Cys) for 72 h. The cells were then stained to identify any senescent cells by a lysosomal β-galactosidase activity assay and p53 and p21 expression. E, F: The percentage of senescent cells in HMDMs (E) and THP-1 cells (F), which had been treated in the same way. The images shown are representative of three independent experiments. G, H: The MFI for p53 (G) and p21 (H) expression in HMDMs. I, J: A comparison of p53 (I) and p21 (J) MFI under various treatment conditions. * P

    Journal: Journal of Lipid Research

    Article Title: Lysosomal oxidation of LDL alters lysosomal pH, induces senescence, and increases secretion of pro-inflammatory cytokines in human macrophages [S]

    doi: 10.1194/jlr.M088245

    Figure Lengend Snippet: Effect of lysosomal oxidation of LDL on senescence in human macrophages. HMDMs were cultured in 12-well tissue culture plates at 3,000 cells per well in RPMI medium (containing 10% v/v lipoprotein-deficient serum) containing either no LDL (A), native LDL (B), SMase-LDL alone (C), or SMase-LDL (D) (all at 100 μg protein/ml) with 10 μM cysteamine (Cys) for 72 h. The cells were then stained to identify any senescent cells by a lysosomal β-galactosidase activity assay and p53 and p21 expression. E, F: The percentage of senescent cells in HMDMs (E) and THP-1 cells (F), which had been treated in the same way. The images shown are representative of three independent experiments. G, H: The MFI for p53 (G) and p21 (H) expression in HMDMs. I, J: A comparison of p53 (I) and p21 (J) MFI under various treatment conditions. * P

    Article Snippet: THP-1 macrophages or HMDMs (4,000 per well) were plated in 12-well tissue culture plates (Corning).

    Techniques: Cell Culture, Staining, Activity Assay, Expressing

    Growth kinetics of Huh7 cell lines. Cells were seeded at 5×10 4 cells/well in a 12-well plate. At indicated times post-seeding, cells were trypsinized and counted. Results are graphed as mean±sem of triplicates.

    Journal: PLoS ONE

    Article Title: Hepatitis C Virus Infection in Phenotypically Distinct Huh7 Cell Lines

    doi: 10.1371/journal.pone.0006561

    Figure Lengend Snippet: Growth kinetics of Huh7 cell lines. Cells were seeded at 5×10 4 cells/well in a 12-well plate. At indicated times post-seeding, cells were trypsinized and counted. Results are graphed as mean±sem of triplicates.

    Article Snippet: Growth kinetics To ensure that none of the Huh7 cell lines would reach confluence during the 3 day period in which growth was monitored, a confluence of ∼40% was targeted by seeding ∼5×104 Huh7 cells in each well of a 12-well tissue culture plate (Corning, Lowell, MA).

    Techniques:

    Morphological analysis of Huh7 cell lines. Huh7 cells were plated at 5×10 4 cells/well in a 12-well plate and photographed 2 days after plating (magnification,×100; inset×400).

    Journal: PLoS ONE

    Article Title: Hepatitis C Virus Infection in Phenotypically Distinct Huh7 Cell Lines

    doi: 10.1371/journal.pone.0006561

    Figure Lengend Snippet: Morphological analysis of Huh7 cell lines. Huh7 cells were plated at 5×10 4 cells/well in a 12-well plate and photographed 2 days after plating (magnification,×100; inset×400).

    Article Snippet: Growth kinetics To ensure that none of the Huh7 cell lines would reach confluence during the 3 day period in which growth was monitored, a confluence of ∼40% was targeted by seeding ∼5×104 Huh7 cells in each well of a 12-well tissue culture plate (Corning, Lowell, MA).

    Techniques:

    Effect of 1,25(OH) 2 D3 on estrogen-induced proliferation of cultured human UF cells. A, HuLM cells (5000 cells/well) in 12-well plates were cultured in phenol free smooth muscle basal medium containing 5% fetal bovine serum. Cells were treated with 10

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: 1,25-Dihydroxyvitamin D3 Regulates Expression of Sex Steroid Receptors in Human Uterine Fibroid Cells

    doi: 10.1210/jc.2014-4011

    Figure Lengend Snippet: Effect of 1,25(OH) 2 D3 on estrogen-induced proliferation of cultured human UF cells. A, HuLM cells (5000 cells/well) in 12-well plates were cultured in phenol free smooth muscle basal medium containing 5% fetal bovine serum. Cells were treated with 10

    Article Snippet: HuLM cells were seeded onto 12-well tissue culture plates (Becton Dickinson) and treated with estrogen in the absence or presence of increasing doses of 1,25(OH)2 D3 at different time points; then cells were counted at each time point as described in the figure legend (see A).

    Techniques: Cell Culture

    Dose-dependent effects of C12 on caspase 3/7 in WT MEF MEF were grown on 12 well plates for two days, and growth medium was changed to Ringer’s solution. C12 was added at the concentrations shown for 1 hr, followed by preparation for measurement of caspase 3/7 activities in RLU. The threshold concentration for C12-triggered activation of caspase was between 1 and 10 μM. Avg +/− SD, n = 3 experiments. * p

    Journal: Cellular microbiology

    Article Title: Pseudomonas aeruginosa Homoserine Lactone Triggers Apoptosis and Bak/Bax-Independent Release of Mitochondrial Cytochrome C in Fibroblasts

    doi: 10.1111/cmi.12263

    Figure Lengend Snippet: Dose-dependent effects of C12 on caspase 3/7 in WT MEF MEF were grown on 12 well plates for two days, and growth medium was changed to Ringer’s solution. C12 was added at the concentrations shown for 1 hr, followed by preparation for measurement of caspase 3/7 activities in RLU. The threshold concentration for C12-triggered activation of caspase was between 1 and 10 μM. Avg +/− SD, n = 3 experiments. * p

    Article Snippet: The cells were passaged at 1:5-1:10 dilutions and the remaining cell suspension was seeded directly onto a 96-well, 24-well or 12-well tissue culture plate (BD Falcon, Bedford, MA) or onto coverglasses for imaging.

    Techniques: Concentration Assay, Activation Assay

    Evaluation of the BBB-penetration ability of CPDs in vitro. (A–C) Schematic diagram of the preparation and application of the in vitro BBB model: (A) the 12-well plates were seeded with C6 cells; (B) HUVEC were seeded into the transwell inserts; (C) CPDs were added into the transwell lumen, and the fluid in the lower compartment containing the CPDs could be detected. (D) Time-dependent TEER values of the in vitro BBB model ( n = 3, mean ± SD). (E) Accumulated percentages of CPDs that crossed the BBB ( n = 6, mean ± SD).

    Journal: ACS Omega

    Article Title: Noninvasive Brain Tumor Imaging Using Red Emissive Carbonized Polymer Dots across the Blood–Brain Barrier

    doi: 10.1021/acsomega.8b01169

    Figure Lengend Snippet: Evaluation of the BBB-penetration ability of CPDs in vitro. (A–C) Schematic diagram of the preparation and application of the in vitro BBB model: (A) the 12-well plates were seeded with C6 cells; (B) HUVEC were seeded into the transwell inserts; (C) CPDs were added into the transwell lumen, and the fluid in the lower compartment containing the CPDs could be detected. (D) Time-dependent TEER values of the in vitro BBB model ( n = 3, mean ± SD). (E) Accumulated percentages of CPDs that crossed the BBB ( n = 6, mean ± SD).

    Article Snippet: First, the polycarbonate membranes of 12-well transwell plates (Corning Incorporated) were coated with rat-tail type I collagen before use.

    Techniques: In Vitro

    hAMCs have no inhibitory effect on THP-1 cells viability. a Cells counting result. PMA-induced THP-1 cells were treated with LPS (10 μg/mL) with or without hAMCs in a 12-well transwell plate for 24 h at 37 °C. The THP-1 cells were then collected and counted. b CCK-8 assay result. THP-1 cells were firstly induced by PMA for 48 h. Then, different concentrations of hAMCs supernatants (0, 20, 40, 80, and 100 %) were added. 20 h later, 10 μL of CCK-8 reagent was added into each well and the cells were incubated for another 4 h. Finally, the absorbance of each well was measured at 450 nm. All experiments were performed three times. Data are the mean ± S.D. of three independent experiments

    Journal: Biological Research

    Article Title: Human amnion mesenchymal cells inhibit lipopolysaccharide-induced TNF-α and IL-1β production in THP-1 cells

    doi: 10.1186/s40659-015-0062-3

    Figure Lengend Snippet: hAMCs have no inhibitory effect on THP-1 cells viability. a Cells counting result. PMA-induced THP-1 cells were treated with LPS (10 μg/mL) with or without hAMCs in a 12-well transwell plate for 24 h at 37 °C. The THP-1 cells were then collected and counted. b CCK-8 assay result. THP-1 cells were firstly induced by PMA for 48 h. Then, different concentrations of hAMCs supernatants (0, 20, 40, 80, and 100 %) were added. 20 h later, 10 μL of CCK-8 reagent was added into each well and the cells were incubated for another 4 h. Finally, the absorbance of each well was measured at 450 nm. All experiments were performed three times. Data are the mean ± S.D. of three independent experiments

    Article Snippet: After washing with PBS three times to remove PMA, they were stimulated with 10 μg/mL LPS (Sigma-Aldrich, St. Louis, MO, USA) with or without hAMCs in a 12-well transwell plate (0.4 μM pore size, Corning, Lowell, MA, USA) for 24 h at 37 °C.

    Techniques: CCK-8 Assay, Incubation

    hAMCs inhibit the phosphorylation of MAPKs and activation of NF-κB in LPS-stimulated THP-1 cells. PMA-induced THP-1 cells were treated with LPS (10 μg/mL) with or without hAMCs (5 × 10 5 , 1 × 10 6 , 2 × 10 6 ) in a 12-well transwell plate for 24 h at 37 °C. The cells were then collected and extracted. Western blot analysis was performed to detect p-ERK, total ERK, p-JNK, total JNK and p65. β-actin was used as an internal control

    Journal: Biological Research

    Article Title: Human amnion mesenchymal cells inhibit lipopolysaccharide-induced TNF-α and IL-1β production in THP-1 cells

    doi: 10.1186/s40659-015-0062-3

    Figure Lengend Snippet: hAMCs inhibit the phosphorylation of MAPKs and activation of NF-κB in LPS-stimulated THP-1 cells. PMA-induced THP-1 cells were treated with LPS (10 μg/mL) with or without hAMCs (5 × 10 5 , 1 × 10 6 , 2 × 10 6 ) in a 12-well transwell plate for 24 h at 37 °C. The cells were then collected and extracted. Western blot analysis was performed to detect p-ERK, total ERK, p-JNK, total JNK and p65. β-actin was used as an internal control

    Article Snippet: After washing with PBS three times to remove PMA, they were stimulated with 10 μg/mL LPS (Sigma-Aldrich, St. Louis, MO, USA) with or without hAMCs in a 12-well transwell plate (0.4 μM pore size, Corning, Lowell, MA, USA) for 24 h at 37 °C.

    Techniques: Activation Assay, Western Blot

    hAMCs co-culture inhibits LPS-induced TNF-α and IL-1β production in THP-1 cells. THP-1 cells were pre-treated with PMA to induce macrophage. Then, they were stimulated with 10 μg/mL LPS with or without different numbers of hAMCs (5 × 10 5 , 1 × 10 6 , 2 × 10 6 ) in a 12-well transwell plate for 24 h at 37 °C. Expression levels of TNF-α ( a ) and IL-1β ( b ) in the culture supernatants were detected by ELISA. Data represent the mean ± S.D. of three independent experiments. **P

    Journal: Biological Research

    Article Title: Human amnion mesenchymal cells inhibit lipopolysaccharide-induced TNF-α and IL-1β production in THP-1 cells

    doi: 10.1186/s40659-015-0062-3

    Figure Lengend Snippet: hAMCs co-culture inhibits LPS-induced TNF-α and IL-1β production in THP-1 cells. THP-1 cells were pre-treated with PMA to induce macrophage. Then, they were stimulated with 10 μg/mL LPS with or without different numbers of hAMCs (5 × 10 5 , 1 × 10 6 , 2 × 10 6 ) in a 12-well transwell plate for 24 h at 37 °C. Expression levels of TNF-α ( a ) and IL-1β ( b ) in the culture supernatants were detected by ELISA. Data represent the mean ± S.D. of three independent experiments. **P

    Article Snippet: After washing with PBS three times to remove PMA, they were stimulated with 10 μg/mL LPS (Sigma-Aldrich, St. Louis, MO, USA) with or without hAMCs in a 12-well transwell plate (0.4 μM pore size, Corning, Lowell, MA, USA) for 24 h at 37 °C.

    Techniques: Co-Culture Assay, Expressing, Enzyme-linked Immunosorbent Assay

    hAMCs co-culture inhibits the mRNA expression of TNF-α and IL-1β in LPS-stimulated THP-1 cells. THP-1 cells were pre-treated with PMA to induce macrophage. Then, they were stimulated with 10 μg/mL LPS with or without different numbers of hAMCs (5 × 10 5 , 1 × 10 6 , 2 × 10 6 ) in a 12-well transwell plate for 24 h at 37 °C. TNF-α mRNA ( a ) and IL-1β mRNA ( b ) expressions were determined by real time-PCR. GAPDH was used as a quantitative control. All experiments were performed three times. **P

    Journal: Biological Research

    Article Title: Human amnion mesenchymal cells inhibit lipopolysaccharide-induced TNF-α and IL-1β production in THP-1 cells

    doi: 10.1186/s40659-015-0062-3

    Figure Lengend Snippet: hAMCs co-culture inhibits the mRNA expression of TNF-α and IL-1β in LPS-stimulated THP-1 cells. THP-1 cells were pre-treated with PMA to induce macrophage. Then, they were stimulated with 10 μg/mL LPS with or without different numbers of hAMCs (5 × 10 5 , 1 × 10 6 , 2 × 10 6 ) in a 12-well transwell plate for 24 h at 37 °C. TNF-α mRNA ( a ) and IL-1β mRNA ( b ) expressions were determined by real time-PCR. GAPDH was used as a quantitative control. All experiments were performed three times. **P

    Article Snippet: After washing with PBS three times to remove PMA, they were stimulated with 10 μg/mL LPS (Sigma-Aldrich, St. Louis, MO, USA) with or without hAMCs in a 12-well transwell plate (0.4 μM pore size, Corning, Lowell, MA, USA) for 24 h at 37 °C.

    Techniques: Co-Culture Assay, Expressing, Real-time Polymerase Chain Reaction

    Formation of syncytia by CC81 cells transfected with infectious BLV clones. CC81 cells were transfected with infectious clones of BLV or the control plasmid. Then, 24 h after transfection, transfected cells were replated at 8 × 10 4 cells/well in 12-well culture plates and cultured for 2 days in complete medium with 4 mg of Polybrene per ml. The cells were fixed and stained, and the syncytia and nuclei in each syncytium were counted. Syncytia were classified into three groups: syncytia with 5 to 9 nuclei (left), 10 to 14 nuclei (center), and 15 or more nuclei (right). Average values from three wells with standard error (error bars) are shown.

    Journal: Journal of Virology

    Article Title: A Mutant Form of the Tax Protein of Bovine Leukemia Virus (BLV), with Enhanced Transactivation Activity, Increases Expression and Propagation of BLV In Vitro but Not In Vivo

    doi: 10.1128/JVI.77.3.1894-1903.2003

    Figure Lengend Snippet: Formation of syncytia by CC81 cells transfected with infectious BLV clones. CC81 cells were transfected with infectious clones of BLV or the control plasmid. Then, 24 h after transfection, transfected cells were replated at 8 × 10 4 cells/well in 12-well culture plates and cultured for 2 days in complete medium with 4 mg of Polybrene per ml. The cells were fixed and stained, and the syncytia and nuclei in each syncytium were counted. Syncytia were classified into three groups: syncytia with 5 to 9 nuclei (left), 10 to 14 nuclei (center), and 15 or more nuclei (right). Average values from three wells with standard error (error bars) are shown.

    Article Snippet: At 24 h after transfection, CC81 cells transfected were replated at 8 × 104 cells/well in 12-well culture plates and cultured for an additional 2 days in RPMI 1640 medium supplemented with 4 mg of Polybrene (Sigma, St. Louis, Mo.) per ml.

    Techniques: Transfection, Clone Assay, Plasmid Preparation, Cell Culture, Staining

    Drawing Voronoi diagrams, and examples of their use. A. Principle. ( i ) In this example (4 points randomly-distributed in a square), ( ii ) we draw straight lines between each point (here shown for point 2) and its nearest neighbors. ( iii ) The corresponding Voronoi polygon is formed using half-plane intersections for all lines. ( iv ) The same procedure is applied for other points to produce the Voronoi diagram. B, C. Example Voronoi diagrams used during cloning of ES and HEK cells. Details are as Fig. 3 , and each pair of images on the left show the same region of the dish immediately after plating cells (d 0; circles surround single cells), and prior to picking colonies (d 7; dotted lines outline colonies). Right-hand panels show views of picked colonies growing in conventional 12-well plates at d 17.

    Journal: bioRxiv

    Article Title: Jet-printing microfluidic devices on demand

    doi: 10.1101/2020.05.31.126300

    Figure Lengend Snippet: Drawing Voronoi diagrams, and examples of their use. A. Principle. ( i ) In this example (4 points randomly-distributed in a square), ( ii ) we draw straight lines between each point (here shown for point 2) and its nearest neighbors. ( iii ) The corresponding Voronoi polygon is formed using half-plane intersections for all lines. ( iv ) The same procedure is applied for other points to produce the Voronoi diagram. B, C. Example Voronoi diagrams used during cloning of ES and HEK cells. Details are as Fig. 3 , and each pair of images on the left show the same region of the dish immediately after plating cells (d 0; circles surround single cells), and prior to picking colonies (d 7; dotted lines outline colonies). Right-hand panels show views of picked colonies growing in conventional 12-well plates at d 17.

    Article Snippet: Retrieved cells were plated in 12-well plates (Greiner Bio-One, Kremsmünster, Austria; #665180) and allowed to reattach and grow.

    Techniques: Clone Assay

    Co-culture model description. IEC were grown in 12-well transwell inserts until confluency and basolaterally exposed to either non-activated or αCD3/CD28-activated PBMC. Apically, IEC were conditioned with 2′-FL or GF in the presence or absence of CpG, a TLR9 agonist mimicking a bacterial trigger ( A ). After 24 h incubation, basolateral supernatant was collected to analyze the T-cell mediator release. The IEC were set apart and washed with PBS. Then, fresh medium was added and IEC were kept in incubation for an additional 24 h to study the IEC-derived mediator release ( B ). Alternatively, IEC were washed with PBS and co-cultured with immature moDC for 48 h ( C ). Then, the basolateral supernatant was collected where the mediator release was studied. Additionally, the phenotype of moDC after IEC/moDC co-culture was analyzed. Subsequently, conditioned moDC (ccDC) were exposed to naïve T-cells in an allogeneic DC/T-cell assay ( D ). After 5–6 days incubation, the cytokine release was measured in the supernatant.

    Journal: Biomolecules

    Article Title: Exposure of Intestinal Epithelial Cells to 2′-Fucosyllactose and CpG Enhances Galectin Release and Instructs Dendritic Cells to Drive Th1 and Regulatory-Type Immune Development

    doi: 10.3390/biom10050784

    Figure Lengend Snippet: Co-culture model description. IEC were grown in 12-well transwell inserts until confluency and basolaterally exposed to either non-activated or αCD3/CD28-activated PBMC. Apically, IEC were conditioned with 2′-FL or GF in the presence or absence of CpG, a TLR9 agonist mimicking a bacterial trigger ( A ). After 24 h incubation, basolateral supernatant was collected to analyze the T-cell mediator release. The IEC were set apart and washed with PBS. Then, fresh medium was added and IEC were kept in incubation for an additional 24 h to study the IEC-derived mediator release ( B ). Alternatively, IEC were washed with PBS and co-cultured with immature moDC for 48 h ( C ). Then, the basolateral supernatant was collected where the mediator release was studied. Additionally, the phenotype of moDC after IEC/moDC co-culture was analyzed. Subsequently, conditioned moDC (ccDC) were exposed to naïve T-cells in an allogeneic DC/T-cell assay ( D ). After 5–6 days incubation, the cytokine release was measured in the supernatant.

    Article Snippet: IEC/PBMC and IEC/moDC Co-Culture Model Description IEC were diluted 5 to 10 times based on surface area and seeded in 12-well transwell inserts (Costar Corning Incorporated, NY, USA) one week prior to the experiments.

    Techniques: Co-Culture Assay, Incubation, Derivative Assay, Cell Culture