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  • 99
    Millipore a549 cells a549 cells
    Effects of MC4 and FK866 on TNFα-induced phosphorylation of NFκB and translocation of NFκB from cytoplasm into the nucleus in <t>A549</t> cells. Notes: A549 cells (3×10 5 ) were starved for 2 h, then pretreated with indicated concentrations of MC4 or FK866 for 2 h, followed by indicated concentrations of MC4 (M) or FK866 (F) together with 50 ng/mL of TNFα treatment for 20 h. ( A ) Western blot analysis of the effects of MC4 and FK866 on TNFα-induced phosphorylation of NFκB. ( B ) Densitometry analysis of TNFα-induced expression of p-NFκB normalized with GAPDH and untreated controls. ( C ) Representative immunofluorescent images showing the effects of MC4 and FK866 on TNFα-stimulated translocation of NFκB. A549 cells were pretreated with 0.3 nM of either MC4 or FK866 for 2 h, then incubated with 20 ng/mL TNFα for 20 min. NFκB antibody was indirectly labeled with Alexa Fluor 488 secondary antibody (green), and cells were mounted with VECTASHIELD Mounting Medium with DAPI (blue). ( D ) Percentage of cells with nuclear translocation of NFκB. A total of 400–500 cells were counted from randomly selected microscope fields of each slide, and the percentage of cells with nuclear translocation of NFκB was calculated. Data are representative of three separate experiments. Bars are mean ± SD, # P
    A549 Cells A549 Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Aviva Systems a549 cells
    Validation of ITGB3 as a direct target of miR-140-3p (A) Predicted binding sites of ITGB3 3’-UTR and miR-140-3p using an online tool TargetScan. The base pairing between the ITGB3 3’UTR binding site and the seed region of miR-140-3p is showed by vertical lines (top). The binding site of miR-140-3p and the mutated form of ITGB3 3’-UTR includes 3 nucleotides (bottom). (B) Relative luciferase activity levels following 24 hours transfection of <t>A549</t> cells with the Luciferase reporter vector containing the backbone vector only, ITGB3 3’-UTR (ITGB3wt) and the mutated form of the 3’-UTR (ITGB3mut), respectively. Data are presented as fold change of the mean + SEM, three individual experiments were undertaken in triplicate, ANOVA-test was used to assess significance with * p
    A549 Cells, supplied by Aviva Systems, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam a549 cells
    Gene mutation and virus replication capability. A: A circos diagram showing the gene mutations. The genome of A/Zhejiang /DTID-ZJU01/2013(H7N9) was used as a reference genome. The circos diagram was constructed to show the mutations in the different genes (HA, NA, PA, PB1, PB2, NP, MA, and NS). In the circos diagram, the innermost circle represents the virus isolated from the plasma. The other circles represent the viruses isolated from the sputum collected at different times. The marked lines show the mutation sites. B-C: The virus replication capability in embryonated chicken eggs. Replication capability was measured by the quantity of RNA and TCID50 titre. D-E: The virus replication capability in embryonated <t>A549</t> cells. Replication capability was measured by the quantity of RNA and TCID50 titre. F: Cell fluorescence map. The virus replication capability was indicated by fluorescence intensity.
    A549 Cells, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 251 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    3-D Matrix a549 cells
    Click-iT Plus EdU proliferation assay. Plot depicts percent of <t>A549</t> cell nuclei positive for EdU on day 4 in monoculture and the A549/CCL-210 co-cultures, separated by gel and media type. The nondegradable gels contained a peptide crosslinker insensitive to MMP cleavage. GM6001 was added at 10 μM, and the DMSO control media contained 0.05% DMSO. The degradable bars refer to the original experiment in MMP-degradable gels with regular growth media. Results are presented as means ± SEM of three biological replicates of each condition. *p
    A549 Cells, supplied by 3-D Matrix, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ibidi a549 cells
    Aspergillus fumigatus hyphae grow parallel to <t>A549</t> cell layer whereas A. niger hyphae grow more perpendicularly. Direction of hyphal growth of A. fumigatus and A. niger in the presence of A549 cells: (A) Z-plane showing thickness of A549 cell layer, nuclei are stained with Hoechst (blue) and cell contour by CellMask TM (green). (B) A549 cell layer X/Y-plane. Z-plane (C,E) and X/Y-plane (D,F) showing A. fumigatus (C,D) and A. niger growth (E,F) on A549 (Hoechst stained) cells. (G) Hyphal growth of A. fumigatus and A. niger in the Z-plane. Bars represent standard deviation. ∗ Indicates significant difference. Approximately 10 fields per slide from three biological replicas were analyzed.
    A549 Cells, supplied by ibidi, used in various techniques. Bioz Stars score: 92/100, based on 176 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Novus Biologicals a549 cells
    NEU3 surface expression on airway ECs. Flow cytometry of surface NEU3 on ( A ) 1HAEo − , ( B ) CFTE29o − , ( C ) 16HBE14o − , ( D ) BEAS-2B, ( E ) SAEC, and ( F ) <t>A549</t> airway ECs. The gray-shaded histograms represent staining with control nonimmune
    A549 Cells, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    DSMZ a549 cells
    Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) <t>A549</t> cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts
    A549 Cells, supplied by DSMZ, used in various techniques. Bioz Stars score: 91/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    National Centre for Cell Science a549 cell lines
    Confocal microscopic images of <t>A549</t> cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.
    A549 Cell Lines, supplied by National Centre for Cell Science, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    LGC Standards GmbH a549 cells
    Differential responses to carboplatin treatment in PC9 and <t>A549</t> cells. A: Carboplatin-induced growth inhibition in PC9 and A549 cells using a sulforhodamine B assay 72 h post treatment. B: Western blot analysis of the levels of uncleaved PARP, cleaved PARP and cleaved (active) caspase 3 72 h post carboplatin treatment (0–200 µM) in PC9 and A549 cells. Actin was used as a loading control. C, D: Flow cytometric analysis of PC9 (C) and A549 cells (D) treated with carboplatin (100 µM) or vehicle. Apoptotic cells were identified by Annexin V-Alexafluor488 (λ Ex/Em = 495/519 nm) and necrotic cells by 7-AAD (λ Ex/Em = 546/647 nm). Population Q4 represents viable cells, whereas population Q3 represents apoptotic cells that have low 7-AAD fluorescence and stain with Annexin V. Population Q2 represents secondary apoptotic/necrotic cells.
    A549 Cells, supplied by LGC Standards GmbH, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Nanjing KeyGen Biotech Co Ltd a549 cells
    ( A ) Antitumor effect of CK and CK mixed micelles to <t>A549</t> cell on different concentrations. * P
    A549 Cells, supplied by Nanjing KeyGen Biotech Co Ltd, used in various techniques. Bioz Stars score: 91/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Genecopoeia a549 cells
    Combinatorial CRISPR screens reveal metabolic network dependencies (A) SKO fitness scores for HeLa cells, plotted as f g (day −1 ), with a more negative score representing a loss in fitness with SKO. Plotted as mean ± SD. (B) Multi-isoform family member fitness scores and gene expression for HeLa (top) and <t>A549</t> (bottom) cells. (C) Relative comparison of SKO fitness scores (f g ) across both cells. (D) Relative comparison of genetic interaction scores (π gg .
    A549 Cells, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 92/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore a549 cells line
    Cell cycle distribution of <t>A549</t> cells line and A549 tumor spheres on effects of chemotherapeutic drugs
    A549 Cells Line, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Diagnostic Hybrids Inc a549 cells
    IFA staining of influenza virus A-infected cells from a clinical specimen. (A) Mv1Lu cells. (B) Mixed <t>A549-Mv1Lu</t> cells. (C) pRhMK cells. Magnification, ×100.
    A549 Cells, supplied by Diagnostic Hybrids Inc, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Viromed a549 cells
    IFA staining of influenza virus A-infected cells from a clinical specimen. (A) Mv1Lu cells. (B) Mixed <t>A549-Mv1Lu</t> cells. (C) pRhMK cells. Magnification, ×100.
    A549 Cells, supplied by Viromed, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher a549 cells
    MTT assays. <t>A549</t> cells were treated with increasing concentration of XAV939 (0.1, 0.5, 1,5 and 10 µM) for 24, 4, 72 and 96 h. The result was shown as relative cell viability per concentration at each time point.
    A549 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 13422 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Microbix a549 cell lines
    MTT assays. <t>A549</t> cells were treated with increasing concentration of XAV939 (0.1, 0.5, 1,5 and 10 µM) for 24, 4, 72 and 96 h. The result was shown as relative cell viability per concentration at each time point.
    A549 Cell Lines, supplied by Microbix, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Corning Life Sciences a549 cells a549 cells
    The effects of calcitriol on <t>A549</t> cells. a Effect of calcitriol treatment on virus growth kinetic in A549 cells. Cell culture supernatants were harvested every 24 h from infected cells and the viral load was titrated using the TCID50 assay. Real-time PCR was used to measure the effect of calcitriol treatment on the mRNA expression levels of the viral M gene ( b ), IL-6 ( c ), and IFN-β ( d ) in A549 cells. β-actin was used as an internal control. The data are expressed as mean ± SEM of triplicate samples
    A549 Cells A549 Cells, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of MC4 and FK866 on TNFα-induced phosphorylation of NFκB and translocation of NFκB from cytoplasm into the nucleus in A549 cells. Notes: A549 cells (3×10 5 ) were starved for 2 h, then pretreated with indicated concentrations of MC4 or FK866 for 2 h, followed by indicated concentrations of MC4 (M) or FK866 (F) together with 50 ng/mL of TNFα treatment for 20 h. ( A ) Western blot analysis of the effects of MC4 and FK866 on TNFα-induced phosphorylation of NFκB. ( B ) Densitometry analysis of TNFα-induced expression of p-NFκB normalized with GAPDH and untreated controls. ( C ) Representative immunofluorescent images showing the effects of MC4 and FK866 on TNFα-stimulated translocation of NFκB. A549 cells were pretreated with 0.3 nM of either MC4 or FK866 for 2 h, then incubated with 20 ng/mL TNFα for 20 min. NFκB antibody was indirectly labeled with Alexa Fluor 488 secondary antibody (green), and cells were mounted with VECTASHIELD Mounting Medium with DAPI (blue). ( D ) Percentage of cells with nuclear translocation of NFκB. A total of 400–500 cells were counted from randomly selected microscope fields of each slide, and the percentage of cells with nuclear translocation of NFκB was calculated. Data are representative of three separate experiments. Bars are mean ± SD, # P

    Journal: Drug Design, Development and Therapy

    Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866

    doi: 10.2147/DDDT.S125349

    Figure Lengend Snippet: Effects of MC4 and FK866 on TNFα-induced phosphorylation of NFκB and translocation of NFκB from cytoplasm into the nucleus in A549 cells. Notes: A549 cells (3×10 5 ) were starved for 2 h, then pretreated with indicated concentrations of MC4 or FK866 for 2 h, followed by indicated concentrations of MC4 (M) or FK866 (F) together with 50 ng/mL of TNFα treatment for 20 h. ( A ) Western blot analysis of the effects of MC4 and FK866 on TNFα-induced phosphorylation of NFκB. ( B ) Densitometry analysis of TNFα-induced expression of p-NFκB normalized with GAPDH and untreated controls. ( C ) Representative immunofluorescent images showing the effects of MC4 and FK866 on TNFα-stimulated translocation of NFκB. A549 cells were pretreated with 0.3 nM of either MC4 or FK866 for 2 h, then incubated with 20 ng/mL TNFα for 20 min. NFκB antibody was indirectly labeled with Alexa Fluor 488 secondary antibody (green), and cells were mounted with VECTASHIELD Mounting Medium with DAPI (blue). ( D ) Percentage of cells with nuclear translocation of NFκB. A total of 400–500 cells were counted from randomly selected microscope fields of each slide, and the percentage of cells with nuclear translocation of NFκB was calculated. Data are representative of three separate experiments. Bars are mean ± SD, # P

    Article Snippet: Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 .

    Techniques: Translocation Assay, Western Blot, Expressing, Incubation, Labeling, Microscopy

    Inhibitory effects of MC4 and FK866 on TNFα-induced intracellular NAD/NADH levels in A549 cells. Notes: A549 cells (3×10 5 ) were starved for 2 h, then pretreated with indicated concentrations of MC4 and FK866 for 2 h, followed by indicated concentrations of MC4 (MC) and FK866 (FK) together with 50 ng/mL of TNFα treatment for 20 h; the intracellular NAD/NADH levels were tested. Results from each group are presented as mean ± SD of three samples from three separate experiments. Bars are mean ± SD, # P

    Journal: Drug Design, Development and Therapy

    Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866

    doi: 10.2147/DDDT.S125349

    Figure Lengend Snippet: Inhibitory effects of MC4 and FK866 on TNFα-induced intracellular NAD/NADH levels in A549 cells. Notes: A549 cells (3×10 5 ) were starved for 2 h, then pretreated with indicated concentrations of MC4 and FK866 for 2 h, followed by indicated concentrations of MC4 (MC) and FK866 (FK) together with 50 ng/mL of TNFα treatment for 20 h; the intracellular NAD/NADH levels were tested. Results from each group are presented as mean ± SD of three samples from three separate experiments. Bars are mean ± SD, # P

    Article Snippet: Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 .

    Techniques:

    Effects of MC4 and FK866 on TNFα-induced cytokine expressions in A549 cells. Notes: After serum-free starvation for 2 h, A549 cells (3×10 5 ) were incubated with indicated concentrations of MC4 and FK866 for 2 h, then treated with MC4 (MC) or FK866 (FK) together with 50 ng/mL TNFα (T) for an additional 5 h. ( A ) A representative gel image of IL6, IL8, IL16, CCR3, NAMPT, and β-actin mRNA levels measured by semi-quantitative RT-PCR. ( B ) Densitometry analysis of TNFα-induced cytokine mRNA levels normalized with β-actin and untreated controls. ( C ) Intracellular IL6 levels were assayed using an intracellular IL6 ELISA kit. Results from each group are presented as mean ± SD of three samples from three separate experiments. # P

    Journal: Drug Design, Development and Therapy

    Article Title: MC-PPEA as a new and more potent inhibitor of CLP-induced sepsis and pulmonary inflammation than FK866

    doi: 10.2147/DDDT.S125349

    Figure Lengend Snippet: Effects of MC4 and FK866 on TNFα-induced cytokine expressions in A549 cells. Notes: After serum-free starvation for 2 h, A549 cells (3×10 5 ) were incubated with indicated concentrations of MC4 and FK866 for 2 h, then treated with MC4 (MC) or FK866 (FK) together with 50 ng/mL TNFα (T) for an additional 5 h. ( A ) A representative gel image of IL6, IL8, IL16, CCR3, NAMPT, and β-actin mRNA levels measured by semi-quantitative RT-PCR. ( B ) Densitometry analysis of TNFα-induced cytokine mRNA levels normalized with β-actin and untreated controls. ( C ) Intracellular IL6 levels were assayed using an intracellular IL6 ELISA kit. Results from each group are presented as mean ± SD of three samples from three separate experiments. # P

    Article Snippet: Immunofluorescence imaging of nuclear translocation of NFκB in A549 cells A549 cells were seeded on poly-L-lysine-coated glass slides (Catalog#: p0425; Sigma-Aldrich) and cultured in a 35 mm Petri dish with 3 mL of media at 37°C in a humidified atmosphere of 5% CO2 .

    Techniques: Incubation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    Entry of rLCMV-LASVGP into A549 epithelial cells shows hallmarks of macropinocytosis. (A) Schema of the inhibitor experiment. For details, see the text. LE, late endosome. (B, C) Entry of rLCMV-LASVGP is independent of dynamin and clathrin. A549 cells

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Entry of rLCMV-LASVGP into A549 epithelial cells shows hallmarks of macropinocytosis. (A) Schema of the inhibitor experiment. For details, see the text. LE, late endosome. (B, C) Entry of rLCMV-LASVGP is independent of dynamin and clathrin. A549 cells

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques:

    Entry of rLCMV-LASVGP does not affect either overall cellular actin dynamics or bulk fluid uptake. (A) Changes in cell morphology during virus entry. Subconfluent A549 cells were mock treated or exposed to rLCMV-LASVGP (100 PFU/cell) or VACV (3 PFU/cell)

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Entry of rLCMV-LASVGP does not affect either overall cellular actin dynamics or bulk fluid uptake. (A) Changes in cell morphology during virus entry. Subconfluent A549 cells were mock treated or exposed to rLCMV-LASVGP (100 PFU/cell) or VACV (3 PFU/cell)

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques:

    Infection of rLCMV -LASVGP in A549 cells depleted of HGFR. (A) Depletion of HGFR by RNAi. A549 cells were transfected with siRNAs specific for HGFR and the corresponding control siRNA (C911). An initial transfection (First) was performed 16 h after plating.

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Infection of rLCMV -LASVGP in A549 cells depleted of HGFR. (A) Depletion of HGFR by RNAi. A549 cells were transfected with siRNAs specific for HGFR and the corresponding control siRNA (C911). An initial transfection (First) was performed 16 h after plating.

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques: Infection, Transfection

    Conserved profile of cellular factors involved in rLCMV-LASVGP entry. (A) Inhibition of rLCMV-LASVGP infection with selected inhibitors in different epithelial cell lines. Monolayers of A549, WI-26VA4, and Caco-2 cells were treated with the indicated

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Conserved profile of cellular factors involved in rLCMV-LASVGP entry. (A) Inhibition of rLCMV-LASVGP infection with selected inhibitors in different epithelial cell lines. Monolayers of A549, WI-26VA4, and Caco-2 cells were treated with the indicated

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques: Inhibition, Infection

    Identification of cellular kinases involved in rLCMV-LASVGP entry. (A) Screening of a library of kinase inhibitors against rLCMV-LASVGP entry. Monolayers of A549 cells were pretreated with inhibitors for 30 min. Infection with rLCMV-LASVGP (MOI = 0.5)

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Identification of cellular kinases involved in rLCMV-LASVGP entry. (A) Screening of a library of kinase inhibitors against rLCMV-LASVGP entry. Monolayers of A549 cells were pretreated with inhibitors for 30 min. Infection with rLCMV-LASVGP (MOI = 0.5)

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques: Infection

    Antiviral activity of the kinase inhibitor EMD 1214063 (EMD). (A) Inhibition of different viruses with EMD 1214063. Monolayers of A549 cells were pretreated with the indicated concentration of EMD 1214063 for 30 min, followed by infection with the different

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Antiviral activity of the kinase inhibitor EMD 1214063 (EMD). (A) Inhibition of different viruses with EMD 1214063. Monolayers of A549 cells were pretreated with the indicated concentration of EMD 1214063 for 30 min, followed by infection with the different

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques: Activity Assay, Inhibition, Concentration Assay, Infection

    Conserved profile of cellular factors involved in rLCMV-LASVGP entry into polarized cells. (A) Detection of functional DG at the apical and basolateral surfaces of polarized A549 and Caco-2 cells. Cells were cultured on Transwell filters to obtain polarized

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Conserved profile of cellular factors involved in rLCMV-LASVGP entry into polarized cells. (A) Detection of functional DG at the apical and basolateral surfaces of polarized A549 and Caco-2 cells. Cells were cultured on Transwell filters to obtain polarized

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques: Functional Assay, Cell Culture

    Additive antiviral activity of EMD 1214063 and ribavirin. (A) Inhibition of rLCMV-LASVGP infection with Rib. A549 cells were infected with rLCMV-LASVGP at an MOI of 0.01. After removal of unbound virus, fresh medium containing the indicated concentrations

    Journal: Journal of Virology

    Article Title: Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis

    doi: 10.1128/JVI.00257-16

    Figure Lengend Snippet: Additive antiviral activity of EMD 1214063 and ribavirin. (A) Inhibition of rLCMV-LASVGP infection with Rib. A549 cells were infected with rLCMV-LASVGP at an MOI of 0.01. After removal of unbound virus, fresh medium containing the indicated concentrations

    Article Snippet: Briefly, A549 cells were serum starved for 16 h and detached with enzyme-free cell dissociation solution (Sigma), and single-cell suspensions (2 × 105 cells/ml) were prepared in PBS containing 2% (wt/vol) bovine serum albumin (BSA).

    Techniques: Activity Assay, Inhibition, Infection

    Histones induce epithelial and endothelial cell death. (A) A549 cells were treated for 16 h with different concentrations of histone type II-A, and the cell morphology was evaluated. (B) A549 cell numbers were counted after treatment with various concentrations of histones for 16 h. (C) HUVEC or A549 cells were treated with 200 µg/ml histones for 16 h or left untreated (control). (D) HUVEC were treated for 16 h with different concentrations of histones, and the extent of cytotoxity was measured. B and D are representative data of three independent experiments, and in A and C pictures are representative pictures from three independent experiments at 20× magnification.

    Journal: PLoS ONE

    Article Title: Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

    doi: 10.1371/journal.pone.0032366

    Figure Lengend Snippet: Histones induce epithelial and endothelial cell death. (A) A549 cells were treated for 16 h with different concentrations of histone type II-A, and the cell morphology was evaluated. (B) A549 cell numbers were counted after treatment with various concentrations of histones for 16 h. (C) HUVEC or A549 cells were treated with 200 µg/ml histones for 16 h or left untreated (control). (D) HUVEC were treated for 16 h with different concentrations of histones, and the extent of cytotoxity was measured. B and D are representative data of three independent experiments, and in A and C pictures are representative pictures from three independent experiments at 20× magnification.

    Article Snippet: Histone treatment of cells A549 cells and HUVEC were treated with different concentrations of histone type IIA from calf (Sigma-Aldrich).

    Techniques:

    NET cause lung epithelial cell death in a concentration-dependent manner. (A) The morphology of A549 cells was evaluated after 4 or 16 h treatment with medium (control), NET or staurosporine. Shown are representative pictures of > 8 independent experiments at 20× magnification. (B) Cell growth from (A) was quantified by measuring the difference between occupied cell area after 16 h and 4 h. (C) Multicaspase activity of A549 cells was measured after 16 h treatment with two concentrations of NET (3.4 and 10.1 µg/ml DNA-NET) or staurosporine. Shown are representative data of three independent experiments (mean SD), * p

    Journal: PLoS ONE

    Article Title: Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

    doi: 10.1371/journal.pone.0032366

    Figure Lengend Snippet: NET cause lung epithelial cell death in a concentration-dependent manner. (A) The morphology of A549 cells was evaluated after 4 or 16 h treatment with medium (control), NET or staurosporine. Shown are representative pictures of > 8 independent experiments at 20× magnification. (B) Cell growth from (A) was quantified by measuring the difference between occupied cell area after 16 h and 4 h. (C) Multicaspase activity of A549 cells was measured after 16 h treatment with two concentrations of NET (3.4 and 10.1 µg/ml DNA-NET) or staurosporine. Shown are representative data of three independent experiments (mean SD), * p

    Article Snippet: Histone treatment of cells A549 cells and HUVEC were treated with different concentrations of histone type IIA from calf (Sigma-Aldrich).

    Techniques: Concentration Assay, Activity Assay

    Inhibition of neutrophil elastase does not inhibit NET-induced cytotoxicity. (A) The supernatants of unstimulated (Unstim) or stimulated (Stim) neutrophils (50 nM PMA for 4 h) were collected and analyzed for elastase activity in the absence ( filled bars ) or presence ( open bars ) of neutrophil elastase inhibitor (NEI). Likewise, NET were isolated from stimulated cells and digested with DNase or MNase or kept undigested (−), followed by analysis of elastase activity in the same way. (B) Cytotoxicity of A549 cells was measured after 16 h treatment with NET (DNase-digested) in the absence or presence of NEI. Similar results were seen for MNase- or non-digested NET as well as with different NEI concentrations from 0.125 to 1 mM. Shown are representative data of three independent experiments (mean SD), *** p

    Journal: PLoS ONE

    Article Title: Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

    doi: 10.1371/journal.pone.0032366

    Figure Lengend Snippet: Inhibition of neutrophil elastase does not inhibit NET-induced cytotoxicity. (A) The supernatants of unstimulated (Unstim) or stimulated (Stim) neutrophils (50 nM PMA for 4 h) were collected and analyzed for elastase activity in the absence ( filled bars ) or presence ( open bars ) of neutrophil elastase inhibitor (NEI). Likewise, NET were isolated from stimulated cells and digested with DNase or MNase or kept undigested (−), followed by analysis of elastase activity in the same way. (B) Cytotoxicity of A549 cells was measured after 16 h treatment with NET (DNase-digested) in the absence or presence of NEI. Similar results were seen for MNase- or non-digested NET as well as with different NEI concentrations from 0.125 to 1 mM. Shown are representative data of three independent experiments (mean SD), *** p

    Article Snippet: Histone treatment of cells A549 cells and HUVEC were treated with different concentrations of histone type IIA from calf (Sigma-Aldrich).

    Techniques: Inhibition, Activity Assay, Isolation

    Myeloperoxidase inhibition moderately decreases NET-induced cytotoxicity of epithelial cells. Nondigested or DNase-digested NET were pre-incubated without or with myeloperoxidase inhibitor (MPOI), followed by incubation of NET with epithelial cells, A549 or AT-II cells, for 16 h and quantification of cytotoxicty. MPOI alone (37 ng/ml) was not toxic for the epithelial cells. Shown are representative data of three (for AT-II cells, n = 2) independent experiments (mean SD), * p

    Journal: PLoS ONE

    Article Title: Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

    doi: 10.1371/journal.pone.0032366

    Figure Lengend Snippet: Myeloperoxidase inhibition moderately decreases NET-induced cytotoxicity of epithelial cells. Nondigested or DNase-digested NET were pre-incubated without or with myeloperoxidase inhibitor (MPOI), followed by incubation of NET with epithelial cells, A549 or AT-II cells, for 16 h and quantification of cytotoxicty. MPOI alone (37 ng/ml) was not toxic for the epithelial cells. Shown are representative data of three (for AT-II cells, n = 2) independent experiments (mean SD), * p

    Article Snippet: Histone treatment of cells A549 cells and HUVEC were treated with different concentrations of histone type IIA from calf (Sigma-Aldrich).

    Techniques: Inhibition, Incubation

    APC decreases epithelial cytotoxicity induced by histones but not by NET. (A) Histones (200 or 100 µg/ml), pre-incubated for 1 h at 37°C in the absence or presence of 100 nM human APC, were incubated with A549 cells for 16 h, followed by analysis of cytotoxicity. (B) NET were incubated with APC (mass ratio APC: NET proteins, 1∶5, 1∶2 and 1∶1) or without APC for 1 h at 37°C, followed by incubation with A549 cells for 16 h and measurement of cytotoxicty. APC alone or active-site blocked APC (APC+PPACK) were incubated with A549 cells for control. (C) DNase-digested and (D) undigested forms of NET were pre-incubated with 100 nM APC for 20 to 80 min before incubation with A549 cells for 16 h, followed by determination of cytotoxicty. Shown are representative data of four independent experiments (mean SD), *** p

    Journal: PLoS ONE

    Article Title: Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

    doi: 10.1371/journal.pone.0032366

    Figure Lengend Snippet: APC decreases epithelial cytotoxicity induced by histones but not by NET. (A) Histones (200 or 100 µg/ml), pre-incubated for 1 h at 37°C in the absence or presence of 100 nM human APC, were incubated with A549 cells for 16 h, followed by analysis of cytotoxicity. (B) NET were incubated with APC (mass ratio APC: NET proteins, 1∶5, 1∶2 and 1∶1) or without APC for 1 h at 37°C, followed by incubation with A549 cells for 16 h and measurement of cytotoxicty. APC alone or active-site blocked APC (APC+PPACK) were incubated with A549 cells for control. (C) DNase-digested and (D) undigested forms of NET were pre-incubated with 100 nM APC for 20 to 80 min before incubation with A549 cells for 16 h, followed by determination of cytotoxicty. Shown are representative data of four independent experiments (mean SD), *** p

    Article Snippet: Histone treatment of cells A549 cells and HUVEC were treated with different concentrations of histone type IIA from calf (Sigma-Aldrich).

    Techniques: Incubation

    Histone antibodies and polysialic acid decrease NET-mediated cytotoxicity. (A) NET were pre-incubated with different antibodies against histones (DNA/H1, H2A, H2B, H3, citrullinated H3 [cit H3], H4) or with isotype-matched control antibodies. Antibody-treated NET or NET alone (−) were incubated with A549 cells for 16 h to analyze the cytotoxicity. Shown are representative data of three independent experiments (mean SD), *** p

    Journal: PLoS ONE

    Article Title: Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

    doi: 10.1371/journal.pone.0032366

    Figure Lengend Snippet: Histone antibodies and polysialic acid decrease NET-mediated cytotoxicity. (A) NET were pre-incubated with different antibodies against histones (DNA/H1, H2A, H2B, H3, citrullinated H3 [cit H3], H4) or with isotype-matched control antibodies. Antibody-treated NET or NET alone (−) were incubated with A549 cells for 16 h to analyze the cytotoxicity. Shown are representative data of three independent experiments (mean SD), *** p

    Article Snippet: Histone treatment of cells A549 cells and HUVEC were treated with different concentrations of histone type IIA from calf (Sigma-Aldrich).

    Techniques: Incubation

    NET induce cytotoxicity in epithelial and endothelial cells independent of digestion. (A) The extent of cytotoxicity was measured after treatment of A549 cells for 16 h with undigested NET (−), completely (DNase), partially digested (MNase) or boiled forms of NET. The same concentration of DNA alone as DNA-NET (3.4 µg/ml) as well as DNase or MNase alone were used as controls. Shown are representative data of five independent experiments (mean SD), *** p

    Journal: PLoS ONE

    Article Title: Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones

    doi: 10.1371/journal.pone.0032366

    Figure Lengend Snippet: NET induce cytotoxicity in epithelial and endothelial cells independent of digestion. (A) The extent of cytotoxicity was measured after treatment of A549 cells for 16 h with undigested NET (−), completely (DNase), partially digested (MNase) or boiled forms of NET. The same concentration of DNA alone as DNA-NET (3.4 µg/ml) as well as DNase or MNase alone were used as controls. Shown are representative data of five independent experiments (mean SD), *** p

    Article Snippet: Histone treatment of cells A549 cells and HUVEC were treated with different concentrations of histone type IIA from calf (Sigma-Aldrich).

    Techniques: Concentration Assay

    miR-505 inhibits cell proliferation and the epithelial-mesenchymal transition process. (A) A reverse transcription-quantitative polymerase chain reaction assay demonstrated the efficiency of miR-505 overexpression and knockdown plasmids in A549 and H460 cells. Cells were transfected with the pri-miR-505 vector or ASO-miR-505 vector and the control groups, respectively. *** P

    Journal: International Journal of Molecular Medicine

    Article Title: miR-505 inhibits cell growth and EMT by targeting MAP3K3 through the AKT-NFκB pathway in NSCLC cells

    doi: 10.3892/ijmm.2018.4041

    Figure Lengend Snippet: miR-505 inhibits cell proliferation and the epithelial-mesenchymal transition process. (A) A reverse transcription-quantitative polymerase chain reaction assay demonstrated the efficiency of miR-505 overexpression and knockdown plasmids in A549 and H460 cells. Cells were transfected with the pri-miR-505 vector or ASO-miR-505 vector and the control groups, respectively. *** P

    Article Snippet: Proliferation assays The cell proliferation was measured in A549 and H460 cells using a Cell Counting Kit-8 (CCK-8; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).

    Techniques: Real-time Polymerase Chain Reaction, Over Expression, Transfection, Plasmid Preparation, Allele-specific Oligonucleotide

    miR-505 inhibits tumor growth in vivo . (A) Compared with the lenti-control group, the tumorigenic ability of A549 cells was inhibited following trans-fection with lenti-miR-505. (B) The tumor growth rate was significantly decreased following treatment with lenti-miR-505. * P

    Journal: International Journal of Molecular Medicine

    Article Title: miR-505 inhibits cell growth and EMT by targeting MAP3K3 through the AKT-NFκB pathway in NSCLC cells

    doi: 10.3892/ijmm.2018.4041

    Figure Lengend Snippet: miR-505 inhibits tumor growth in vivo . (A) Compared with the lenti-control group, the tumorigenic ability of A549 cells was inhibited following trans-fection with lenti-miR-505. (B) The tumor growth rate was significantly decreased following treatment with lenti-miR-505. * P

    Article Snippet: Proliferation assays The cell proliferation was measured in A549 and H460 cells using a Cell Counting Kit-8 (CCK-8; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).

    Techniques: In Vivo

    MAP3K3 promotes cell proliferation and the epithelial-mesenchymal transition process. (A) A reverse transcription-quantitative polymerase chain reaction assay demonstrated the efficiency of the overexpression plasmid of MAP3K3 in A549 and H460 cells. (B) The indicated transfection on A549 and H460 cellular viabilities was determined with a Cell Counting Kit-8 assay. A549 and H460 cells were transfected with the indicated combinations of pcDNA3 and MAP3K3 or pri-miR-505 and MAP3K3 or the control group. MAP3K3 overexpression promoted cell viability. NS vs. pri-miR-505 and MAP3K3; ** P

    Journal: International Journal of Molecular Medicine

    Article Title: miR-505 inhibits cell growth and EMT by targeting MAP3K3 through the AKT-NFκB pathway in NSCLC cells

    doi: 10.3892/ijmm.2018.4041

    Figure Lengend Snippet: MAP3K3 promotes cell proliferation and the epithelial-mesenchymal transition process. (A) A reverse transcription-quantitative polymerase chain reaction assay demonstrated the efficiency of the overexpression plasmid of MAP3K3 in A549 and H460 cells. (B) The indicated transfection on A549 and H460 cellular viabilities was determined with a Cell Counting Kit-8 assay. A549 and H460 cells were transfected with the indicated combinations of pcDNA3 and MAP3K3 or pri-miR-505 and MAP3K3 or the control group. MAP3K3 overexpression promoted cell viability. NS vs. pri-miR-505 and MAP3K3; ** P

    Article Snippet: Proliferation assays The cell proliferation was measured in A549 and H460 cells using a Cell Counting Kit-8 (CCK-8; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).

    Techniques: Real-time Polymerase Chain Reaction, Over Expression, Plasmid Preparation, Transfection, Cell Counting

    miR-505 inhibits the AKT-nuclear factor-κB pathway through MAP3K3. (A) A western blot assay demonstrated the protein expression levels of IKKa, IKKβ, pAKT, AKT with pri-miR-505 or ASO-miR-505 and the control group in A549 cells. miR-505 overexpression inhibited IKKa, IKKβ and pAKT expression but not the total AKT expression. ** P

    Journal: International Journal of Molecular Medicine

    Article Title: miR-505 inhibits cell growth and EMT by targeting MAP3K3 through the AKT-NFκB pathway in NSCLC cells

    doi: 10.3892/ijmm.2018.4041

    Figure Lengend Snippet: miR-505 inhibits the AKT-nuclear factor-κB pathway through MAP3K3. (A) A western blot assay demonstrated the protein expression levels of IKKa, IKKβ, pAKT, AKT with pri-miR-505 or ASO-miR-505 and the control group in A549 cells. miR-505 overexpression inhibited IKKa, IKKβ and pAKT expression but not the total AKT expression. ** P

    Article Snippet: Proliferation assays The cell proliferation was measured in A549 and H460 cells using a Cell Counting Kit-8 (CCK-8; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).

    Techniques: Western Blot, Expressing, Allele-specific Oligonucleotide, Over Expression

    miR-505 directly targets MAP3K3. (A) The predicted miR-505 binding sites in MAP3K3 mRNA using microRNA.org were depicted. (B) The sequence of MAP3K3-3′UTR-Mut was depicted. (C) A549 and H460 cells were co-transfected with pcDNA3/EGFP-MAP3K3 3′-UTR or 3′-UTR-mut and pri-miR-505 or ASO-miR-505 and the control group. The EGFP intensity was determined with a spectrophotometer, and the value of the control group (pcDNA3 or ASO-NC) was set to 1. *** P

    Journal: International Journal of Molecular Medicine

    Article Title: miR-505 inhibits cell growth and EMT by targeting MAP3K3 through the AKT-NFκB pathway in NSCLC cells

    doi: 10.3892/ijmm.2018.4041

    Figure Lengend Snippet: miR-505 directly targets MAP3K3. (A) The predicted miR-505 binding sites in MAP3K3 mRNA using microRNA.org were depicted. (B) The sequence of MAP3K3-3′UTR-Mut was depicted. (C) A549 and H460 cells were co-transfected with pcDNA3/EGFP-MAP3K3 3′-UTR or 3′-UTR-mut and pri-miR-505 or ASO-miR-505 and the control group. The EGFP intensity was determined with a spectrophotometer, and the value of the control group (pcDNA3 or ASO-NC) was set to 1. *** P

    Article Snippet: Proliferation assays The cell proliferation was measured in A549 and H460 cells using a Cell Counting Kit-8 (CCK-8; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).

    Techniques: Binding Assay, Sequencing, Transfection, Allele-specific Oligonucleotide, Spectrophotometry

    Treatment with TSA inhibits CUG2-induced cell migration, invasion and sphere formation. (A) A549-CUG2 cells were treated with TSA (100 nM) or DMSO for 24 h, and the cell monolayer was scratched. Cell migration was measured by a wound healing assay. (B) An invasion assay was performed after treatment with TSA or DMSO using 48-well Boyden chambers coated with Matrigel. Scale bar indicates 100 µm (***P

    Journal: Oncology Reports

    Article Title: STAT1-HDAC4 signaling induces epithelial-mesenchymal transition and sphere formation of cancer cells overexpressing the oncogene, CUG2

    doi: 10.3892/or.2018.6701

    Figure Lengend Snippet: Treatment with TSA inhibits CUG2-induced cell migration, invasion and sphere formation. (A) A549-CUG2 cells were treated with TSA (100 nM) or DMSO for 24 h, and the cell monolayer was scratched. Cell migration was measured by a wound healing assay. (B) An invasion assay was performed after treatment with TSA or DMSO using 48-well Boyden chambers coated with Matrigel. Scale bar indicates 100 µm (***P

    Article Snippet: Cell cultures Human lung cancer A549 cells (ATCC, Manassas, VA, USA) stably expressing the vector alone (A549-Vec) or CUG2 (A549-CUG2), and A549-CUG2 cells with stably silenced STAT1 (A549-CUG2-shSTAT1) or the control (A549-CUG2-shVec) were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin, 1% streptomycin, and G418 (0.5 mg/ml; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) at 37°C with 5% CO2 .

    Techniques: Migration, Wound Healing Assay, Invasion Assay

    Constitutive suppression of STAT1 inhibits CUG2-induced cell migration, invasion, and sphere formation. (A) After transfection with sh-STAT1 (A549-CUG2-shSTAT1) or control plasmid (A549-CUG2-shVec) and selection under puromycin (1 µg/ml), suppression of STAT1 expression was confirmed by immunoblotting using an anti-STAT1 antibody. (B) After confluence of A549-CUG2-shSTAT1 and A549-CUG2-shVec cells, the cell monolayer was scratched. Cell migration was measured by a wound healing assay. (C) An invasion assay was compared between A549-CUG2-shSTAT1 and A549-CUG2-shVec cells using 48-well Boyden chambers coated with Matrigel. Scale bar indicates 100 µm (***P

    Journal: Oncology Reports

    Article Title: STAT1-HDAC4 signaling induces epithelial-mesenchymal transition and sphere formation of cancer cells overexpressing the oncogene, CUG2

    doi: 10.3892/or.2018.6701

    Figure Lengend Snippet: Constitutive suppression of STAT1 inhibits CUG2-induced cell migration, invasion, and sphere formation. (A) After transfection with sh-STAT1 (A549-CUG2-shSTAT1) or control plasmid (A549-CUG2-shVec) and selection under puromycin (1 µg/ml), suppression of STAT1 expression was confirmed by immunoblotting using an anti-STAT1 antibody. (B) After confluence of A549-CUG2-shSTAT1 and A549-CUG2-shVec cells, the cell monolayer was scratched. Cell migration was measured by a wound healing assay. (C) An invasion assay was compared between A549-CUG2-shSTAT1 and A549-CUG2-shVec cells using 48-well Boyden chambers coated with Matrigel. Scale bar indicates 100 µm (***P

    Article Snippet: Cell cultures Human lung cancer A549 cells (ATCC, Manassas, VA, USA) stably expressing the vector alone (A549-Vec) or CUG2 (A549-CUG2), and A549-CUG2 cells with stably silenced STAT1 (A549-CUG2-shSTAT1) or the control (A549-CUG2-shVec) were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin, 1% streptomycin, and G418 (0.5 mg/ml; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) at 37°C with 5% CO2 .

    Techniques: Migration, Transfection, Plasmid Preparation, Selection, Expressing, Wound Healing Assay, Invasion Assay

    Validation of ITGB3 as a direct target of miR-140-3p (A) Predicted binding sites of ITGB3 3’-UTR and miR-140-3p using an online tool TargetScan. The base pairing between the ITGB3 3’UTR binding site and the seed region of miR-140-3p is showed by vertical lines (top). The binding site of miR-140-3p and the mutated form of ITGB3 3’-UTR includes 3 nucleotides (bottom). (B) Relative luciferase activity levels following 24 hours transfection of A549 cells with the Luciferase reporter vector containing the backbone vector only, ITGB3 3’-UTR (ITGB3wt) and the mutated form of the 3’-UTR (ITGB3mut), respectively. Data are presented as fold change of the mean + SEM, three individual experiments were undertaken in triplicate, ANOVA-test was used to assess significance with * p

    Journal: Oncotarget

    Article Title: Distinct mechanisms by which two forms of miR-140 suppress the malignant properties of lung cancer cells

    doi: 10.18632/oncotarget.26356

    Figure Lengend Snippet: Validation of ITGB3 as a direct target of miR-140-3p (A) Predicted binding sites of ITGB3 3’-UTR and miR-140-3p using an online tool TargetScan. The base pairing between the ITGB3 3’UTR binding site and the seed region of miR-140-3p is showed by vertical lines (top). The binding site of miR-140-3p and the mutated form of ITGB3 3’-UTR includes 3 nucleotides (bottom). (B) Relative luciferase activity levels following 24 hours transfection of A549 cells with the Luciferase reporter vector containing the backbone vector only, ITGB3 3’-UTR (ITGB3wt) and the mutated form of the 3’-UTR (ITGB3mut), respectively. Data are presented as fold change of the mean + SEM, three individual experiments were undertaken in triplicate, ANOVA-test was used to assess significance with * p

    Article Snippet: The human vascular endothelial growth factor A (VEGF-A) levels in the TCM of A549 cells transfected with negative and miR-140-3p mimics were quantified by using 96-well plates pre-coated with VEGF-A primary antibody by using a specific kit purchased from Aviva System Biology, San Diego, USA.

    Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Plasmid Preparation

    Change of miR expression and invasion properties in lung cancer cells after transfection with miR-140-3p mimic (A) Expression of miR-140-3p in A549 cells. (B) Expression of miR-140-3p in SK-MES-1 cells. (C) Transwell invasion property of A549 cells with response to miR-130-3p mimic. (D) Transwell invasion property of SK-MES-1 cells with response to miR-130-3p mimic. (E) Transwell migration property of A549 cells with response to miR-130-3p mimic. (F) Transwell migration property of SK-MES-1 cells with response to miR-130-3p mimic. Data are presented as mean + SEM, three individual experiments were undertaken in triplicate, Student's t -test was used to assess significance.

    Journal: Oncotarget

    Article Title: Distinct mechanisms by which two forms of miR-140 suppress the malignant properties of lung cancer cells

    doi: 10.18632/oncotarget.26356

    Figure Lengend Snippet: Change of miR expression and invasion properties in lung cancer cells after transfection with miR-140-3p mimic (A) Expression of miR-140-3p in A549 cells. (B) Expression of miR-140-3p in SK-MES-1 cells. (C) Transwell invasion property of A549 cells with response to miR-130-3p mimic. (D) Transwell invasion property of SK-MES-1 cells with response to miR-130-3p mimic. (E) Transwell migration property of A549 cells with response to miR-130-3p mimic. (F) Transwell migration property of SK-MES-1 cells with response to miR-130-3p mimic. Data are presented as mean + SEM, three individual experiments were undertaken in triplicate, Student's t -test was used to assess significance.

    Article Snippet: The human vascular endothelial growth factor A (VEGF-A) levels in the TCM of A549 cells transfected with negative and miR-140-3p mimics were quantified by using 96-well plates pre-coated with VEGF-A primary antibody by using a specific kit purchased from Aviva System Biology, San Diego, USA.

    Techniques: Expressing, Transfection, Migration

    The two strands of miR-140 have minor impact upon the metabolite profile of lung cancer cells Lung cancer cells (A549 and SK-MES-1) and BEAS-2B lung epithelial cells were treated with miRNA mimics for 24 hours before cell samples were collected for metabolite analysis using Mass spectrometry (GC-MS). (A) A principle component analysis of the metabolite data set for the three cell lines without treatment (NEG) and with either miRNA mimic. (B) Example GCMS chromatogram of m/z 174 representing glycine levels in the control (NEG, black line) and after treatment with miR-140-3P (red line) and miR-140-5P (blue line). (C) Normalized peak area of metabolites which differed in level in A549 cells dependent on treatment. (D) Normalized peak area of metabolites which differed in level dependent on the cell line studied. Data are presented as mean + SEM, experiments were undertaken in triplicate, T -test and Mann Whitney tests were used to assess significance among multiple groups with * p

    Journal: Oncotarget

    Article Title: Distinct mechanisms by which two forms of miR-140 suppress the malignant properties of lung cancer cells

    doi: 10.18632/oncotarget.26356

    Figure Lengend Snippet: The two strands of miR-140 have minor impact upon the metabolite profile of lung cancer cells Lung cancer cells (A549 and SK-MES-1) and BEAS-2B lung epithelial cells were treated with miRNA mimics for 24 hours before cell samples were collected for metabolite analysis using Mass spectrometry (GC-MS). (A) A principle component analysis of the metabolite data set for the three cell lines without treatment (NEG) and with either miRNA mimic. (B) Example GCMS chromatogram of m/z 174 representing glycine levels in the control (NEG, black line) and after treatment with miR-140-3P (red line) and miR-140-5P (blue line). (C) Normalized peak area of metabolites which differed in level in A549 cells dependent on treatment. (D) Normalized peak area of metabolites which differed in level dependent on the cell line studied. Data are presented as mean + SEM, experiments were undertaken in triplicate, T -test and Mann Whitney tests were used to assess significance among multiple groups with * p

    Article Snippet: The human vascular endothelial growth factor A (VEGF-A) levels in the TCM of A549 cells transfected with negative and miR-140-3p mimics were quantified by using 96-well plates pre-coated with VEGF-A primary antibody by using a specific kit purchased from Aviva System Biology, San Diego, USA.

    Techniques: Mass Spectrometry, Gas Chromatography-Mass Spectrometry, MANN-WHITNEY

    Tubule formation ability of the primary endothelial cells HUVECs co-cultured with TCM of lung cancer cells treated with miR-140-3p and 140-5p mimics HUVECs were seeded onto a layer of Matrigel with completed growth medium supplemented with 40% TCM from miR-treated lung cancer cells for 24 and 48 hours, respectively. The tubule formation was monitored in real-time by using the EVOS Cell Imaging System. Images were taken every 30 minutes for 6 hours. (A) Total length of the tubule structure form by HUVECs in the presence of TCM from A549. (B) Number of closed mesh structure formed by HUVECs in the presence of TCM from A549. (C) Total length of the tubule structure form by HUVECs in the presence of TCM from SK-MES-1. (D) Number of closed mesh structure formed by HUVECs in the presence of TCM from SK-MES-1. Data are presented as mean + SEM, three individual experiments were undertaken in triplicate, Kruskal-Wallis-test was used to assess significance with * p

    Journal: Oncotarget

    Article Title: Distinct mechanisms by which two forms of miR-140 suppress the malignant properties of lung cancer cells

    doi: 10.18632/oncotarget.26356

    Figure Lengend Snippet: Tubule formation ability of the primary endothelial cells HUVECs co-cultured with TCM of lung cancer cells treated with miR-140-3p and 140-5p mimics HUVECs were seeded onto a layer of Matrigel with completed growth medium supplemented with 40% TCM from miR-treated lung cancer cells for 24 and 48 hours, respectively. The tubule formation was monitored in real-time by using the EVOS Cell Imaging System. Images were taken every 30 minutes for 6 hours. (A) Total length of the tubule structure form by HUVECs in the presence of TCM from A549. (B) Number of closed mesh structure formed by HUVECs in the presence of TCM from A549. (C) Total length of the tubule structure form by HUVECs in the presence of TCM from SK-MES-1. (D) Number of closed mesh structure formed by HUVECs in the presence of TCM from SK-MES-1. Data are presented as mean + SEM, three individual experiments were undertaken in triplicate, Kruskal-Wallis-test was used to assess significance with * p

    Article Snippet: The human vascular endothelial growth factor A (VEGF-A) levels in the TCM of A549 cells transfected with negative and miR-140-3p mimics were quantified by using 96-well plates pre-coated with VEGF-A primary antibody by using a specific kit purchased from Aviva System Biology, San Diego, USA.

    Techniques: Cell Culture, Imaging

    Gene mutation and virus replication capability. A: A circos diagram showing the gene mutations. The genome of A/Zhejiang /DTID-ZJU01/2013(H7N9) was used as a reference genome. The circos diagram was constructed to show the mutations in the different genes (HA, NA, PA, PB1, PB2, NP, MA, and NS). In the circos diagram, the innermost circle represents the virus isolated from the plasma. The other circles represent the viruses isolated from the sputum collected at different times. The marked lines show the mutation sites. B-C: The virus replication capability in embryonated chicken eggs. Replication capability was measured by the quantity of RNA and TCID50 titre. D-E: The virus replication capability in embryonated A549 cells. Replication capability was measured by the quantity of RNA and TCID50 titre. F: Cell fluorescence map. The virus replication capability was indicated by fluorescence intensity.

    Journal: Emerging Microbes & Infections

    Article Title: Novel pathogenic characteristics of highly pathogenic avian influenza virus H7N9: viraemia and extrapulmonary infection

    doi: 10.1080/22221751.2020.1754135

    Figure Lengend Snippet: Gene mutation and virus replication capability. A: A circos diagram showing the gene mutations. The genome of A/Zhejiang /DTID-ZJU01/2013(H7N9) was used as a reference genome. The circos diagram was constructed to show the mutations in the different genes (HA, NA, PA, PB1, PB2, NP, MA, and NS). In the circos diagram, the innermost circle represents the virus isolated from the plasma. The other circles represent the viruses isolated from the sputum collected at different times. The marked lines show the mutation sites. B-C: The virus replication capability in embryonated chicken eggs. Replication capability was measured by the quantity of RNA and TCID50 titre. D-E: The virus replication capability in embryonated A549 cells. Replication capability was measured by the quantity of RNA and TCID50 titre. F: Cell fluorescence map. The virus replication capability was indicated by fluorescence intensity.

    Article Snippet: A549 cells were cultured in DMEM supplemented with 10% FBS at 37°C and 5% CO2.

    Techniques: Mutagenesis, Construct, Isolation, Fluorescence

    Isolation of exosomes and identification of viral genes. A: The culture supernatant from A549 cell model without virus infection was observed using transmission electronic microscope. The exosomes were showed by blue arrow. B: The culture supernatant from A549 cell model with ZJU01 strain infection was observed using transmission electronic microscope. The exosome was showed by blue arrow. The viral particles were viewed and showed by red arrow. C: The culture supernatant from A549 cell model with GZ8H002 strain infection was observed using transmission electronic microscope. The exosome was showed by blue arrow. The viral particles were viewed and showed by red arrow. D: The viral content in A549 cell model infected with ZJU01 strain. E: The viral content in A549 cell model infected with GZ8H002 strain. F: The H7 gene content in exosomes. G: The isolation of H7N9 virus from exosomes. H: The TCID50 titre of the H7N9 virus isolated from the exosomes.

    Journal: Emerging Microbes & Infections

    Article Title: Novel pathogenic characteristics of highly pathogenic avian influenza virus H7N9: viraemia and extrapulmonary infection

    doi: 10.1080/22221751.2020.1754135

    Figure Lengend Snippet: Isolation of exosomes and identification of viral genes. A: The culture supernatant from A549 cell model without virus infection was observed using transmission electronic microscope. The exosomes were showed by blue arrow. B: The culture supernatant from A549 cell model with ZJU01 strain infection was observed using transmission electronic microscope. The exosome was showed by blue arrow. The viral particles were viewed and showed by red arrow. C: The culture supernatant from A549 cell model with GZ8H002 strain infection was observed using transmission electronic microscope. The exosome was showed by blue arrow. The viral particles were viewed and showed by red arrow. D: The viral content in A549 cell model infected with ZJU01 strain. E: The viral content in A549 cell model infected with GZ8H002 strain. F: The H7 gene content in exosomes. G: The isolation of H7N9 virus from exosomes. H: The TCID50 titre of the H7N9 virus isolated from the exosomes.

    Article Snippet: A549 cells were cultured in DMEM supplemented with 10% FBS at 37°C and 5% CO2.

    Techniques: Isolation, Infection, Transmission Assay, Microscopy

    Click-iT Plus EdU proliferation assay. Plot depicts percent of A549 cell nuclei positive for EdU on day 4 in monoculture and the A549/CCL-210 co-cultures, separated by gel and media type. The nondegradable gels contained a peptide crosslinker insensitive to MMP cleavage. GM6001 was added at 10 μM, and the DMSO control media contained 0.05% DMSO. The degradable bars refer to the original experiment in MMP-degradable gels with regular growth media. Results are presented as means ± SEM of three biological replicates of each condition. *p

    Journal: Biomaterials

    Article Title: Epithelial-mesenchymal crosstalk influences cellular behavior in a 3D alveolus-fibroblast model system

    doi: 10.1016/j.biomaterials.2017.11.008

    Figure Lengend Snippet: Click-iT Plus EdU proliferation assay. Plot depicts percent of A549 cell nuclei positive for EdU on day 4 in monoculture and the A549/CCL-210 co-cultures, separated by gel and media type. The nondegradable gels contained a peptide crosslinker insensitive to MMP cleavage. GM6001 was added at 10 μM, and the DMSO control media contained 0.05% DMSO. The degradable bars refer to the original experiment in MMP-degradable gels with regular growth media. Results are presented as means ± SEM of three biological replicates of each condition. *p

    Article Snippet: Furthermore, this inhibitor has previously been used with A549 cells to effectively block invasion into a 3D matrix and has been shown to reduce the fluorescence signal from the MMP sensor peptide used in this work [ , , ].

    Techniques: Proliferation Assay

    Aspergillus fumigatus hyphae grow parallel to A549 cell layer whereas A. niger hyphae grow more perpendicularly. Direction of hyphal growth of A. fumigatus and A. niger in the presence of A549 cells: (A) Z-plane showing thickness of A549 cell layer, nuclei are stained with Hoechst (blue) and cell contour by CellMask TM (green). (B) A549 cell layer X/Y-plane. Z-plane (C,E) and X/Y-plane (D,F) showing A. fumigatus (C,D) and A. niger growth (E,F) on A549 (Hoechst stained) cells. (G) Hyphal growth of A. fumigatus and A. niger in the Z-plane. Bars represent standard deviation. ∗ Indicates significant difference. Approximately 10 fields per slide from three biological replicas were analyzed.

    Journal: Frontiers in Microbiology

    Article Title: Hide, Keep Quiet, and Keep Low: Properties That Make Aspergillus fumigatus a Successful Lung Pathogen

    doi: 10.3389/fmicb.2016.00438

    Figure Lengend Snippet: Aspergillus fumigatus hyphae grow parallel to A549 cell layer whereas A. niger hyphae grow more perpendicularly. Direction of hyphal growth of A. fumigatus and A. niger in the presence of A549 cells: (A) Z-plane showing thickness of A549 cell layer, nuclei are stained with Hoechst (blue) and cell contour by CellMask TM (green). (B) A549 cell layer X/Y-plane. Z-plane (C,E) and X/Y-plane (D,F) showing A. fumigatus (C,D) and A. niger growth (E,F) on A549 (Hoechst stained) cells. (G) Hyphal growth of A. fumigatus and A. niger in the Z-plane. Bars represent standard deviation. ∗ Indicates significant difference. Approximately 10 fields per slide from three biological replicas were analyzed.

    Article Snippet: Germination and Directionality of Hyphal Growth upon Interaction with A549 Cells For germination experiments A549 cells were grown on 8-mm glass coverslips (WPI international BV, Europe) and in μ-slide eight well chambers (Ibidi® , Munich, Germany) for observing hyphal growth directionality.

    Techniques: Staining, Standard Deviation

    Germination and hyphal length of A. fumigatus are more effectively decreased in the presence of A549 cells than that of A. niger . (A) Germination and (B) hyphal length. Bar represents standard error of the mean. ∗ Indicates significant difference. Data are obtained from three separate experiments; at least 100 conidia per condition were analyzed.

    Journal: Frontiers in Microbiology

    Article Title: Hide, Keep Quiet, and Keep Low: Properties That Make Aspergillus fumigatus a Successful Lung Pathogen

    doi: 10.3389/fmicb.2016.00438

    Figure Lengend Snippet: Germination and hyphal length of A. fumigatus are more effectively decreased in the presence of A549 cells than that of A. niger . (A) Germination and (B) hyphal length. Bar represents standard error of the mean. ∗ Indicates significant difference. Data are obtained from three separate experiments; at least 100 conidia per condition were analyzed.

    Article Snippet: Germination and Directionality of Hyphal Growth upon Interaction with A549 Cells For germination experiments A549 cells were grown on 8-mm glass coverslips (WPI international BV, Europe) and in μ-slide eight well chambers (Ibidi® , Munich, Germany) for observing hyphal growth directionality.

    Techniques:

    Polymorphonuclear neutrophils reduce A. niger germination and hyphal length at the surface of A549 cells. A. fumigatus and A. niger percentage of germination (A,C) and hyphal length (B,D) in the presence of A549 cells. Bars represent standard error of the mean. ∗ Indicate significant differences. Data are obtained from three separate experiments; at least 100 conidia per condition were analyzed.

    Journal: Frontiers in Microbiology

    Article Title: Hide, Keep Quiet, and Keep Low: Properties That Make Aspergillus fumigatus a Successful Lung Pathogen

    doi: 10.3389/fmicb.2016.00438

    Figure Lengend Snippet: Polymorphonuclear neutrophils reduce A. niger germination and hyphal length at the surface of A549 cells. A. fumigatus and A. niger percentage of germination (A,C) and hyphal length (B,D) in the presence of A549 cells. Bars represent standard error of the mean. ∗ Indicate significant differences. Data are obtained from three separate experiments; at least 100 conidia per condition were analyzed.

    Article Snippet: Germination and Directionality of Hyphal Growth upon Interaction with A549 Cells For germination experiments A549 cells were grown on 8-mm glass coverslips (WPI international BV, Europe) and in μ-slide eight well chambers (Ibidi® , Munich, Germany) for observing hyphal growth directionality.

    Techniques:

    Cytochalasin-B and nocodazole block internalization of Aspergillus fumigatus more effectively than that of A. niger . Internalization of A. niger (AN) and A. fumigatus (AF) by A549 after treatment with 10 μM cytochalasin-B (CB), and/or 20 μM nocodazole (Noc). For analysis, each conidium was scored as either inside or outside the epithelial cells. A chi-square proportion test was performed using a z -test (α = 0.01) and adjusting P -values for multiple comparisons using the Bonferroni correction method. ∗ Indicates significant difference.

    Journal: Frontiers in Microbiology

    Article Title: Hide, Keep Quiet, and Keep Low: Properties That Make Aspergillus fumigatus a Successful Lung Pathogen

    doi: 10.3389/fmicb.2016.00438

    Figure Lengend Snippet: Cytochalasin-B and nocodazole block internalization of Aspergillus fumigatus more effectively than that of A. niger . Internalization of A. niger (AN) and A. fumigatus (AF) by A549 after treatment with 10 μM cytochalasin-B (CB), and/or 20 μM nocodazole (Noc). For analysis, each conidium was scored as either inside or outside the epithelial cells. A chi-square proportion test was performed using a z -test (α = 0.01) and adjusting P -values for multiple comparisons using the Bonferroni correction method. ∗ Indicates significant difference.

    Article Snippet: Germination and Directionality of Hyphal Growth upon Interaction with A549 Cells For germination experiments A549 cells were grown on 8-mm glass coverslips (WPI international BV, Europe) and in μ-slide eight well chambers (Ibidi® , Munich, Germany) for observing hyphal growth directionality.

    Techniques: Blocking Assay

    NEU3 surface expression on airway ECs. Flow cytometry of surface NEU3 on ( A ) 1HAEo − , ( B ) CFTE29o − , ( C ) 16HBE14o − , ( D ) BEAS-2B, ( E ) SAEC, and ( F ) A549 airway ECs. The gray-shaded histograms represent staining with control nonimmune

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Human airway epithelia express catalytically active NEU3 sialidase

    doi: 10.1152/ajplung.00322.2013

    Figure Lengend Snippet: NEU3 surface expression on airway ECs. Flow cytometry of surface NEU3 on ( A ) 1HAEo − , ( B ) CFTE29o − , ( C ) 16HBE14o − , ( D ) BEAS-2B, ( E ) SAEC, and ( F ) A549 airway ECs. The gray-shaded histograms represent staining with control nonimmune

    Article Snippet: Nonpermeabilized unstimulated 1HAEo− , CFTE29o− , 16HBE14o− , and BEAS-2B cells, SAECs, and A549 cells were stained with rabbit polyclonal anti-human NEU3 antibody (Novus Biologicals, Littleton, CO) or control nonimmune rabbit IgG followed by phycoerythrin-conjugated goat anti-rabbit secondary antibody (Jackson ImmunoResearch).

    Techniques: Expressing, Flow Cytometry, Cytometry, Staining

    Airway epithelial cell (EC) sialidase activity. A : increasing primary small airway EC (SAEC; n = 2) and A549 ( n = 3) cell numbers were assayed for sialidase activity for a ganglioside mixture. B : A549 cells (1.0 × 10 6 ) were assayed for sialidase

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Human airway epithelia express catalytically active NEU3 sialidase

    doi: 10.1152/ajplung.00322.2013

    Figure Lengend Snippet: Airway epithelial cell (EC) sialidase activity. A : increasing primary small airway EC (SAEC; n = 2) and A549 ( n = 3) cell numbers were assayed for sialidase activity for a ganglioside mixture. B : A549 cells (1.0 × 10 6 ) were assayed for sialidase

    Article Snippet: Nonpermeabilized unstimulated 1HAEo− , CFTE29o− , 16HBE14o− , and BEAS-2B cells, SAECs, and A549 cells were stained with rabbit polyclonal anti-human NEU3 antibody (Novus Biologicals, Littleton, CO) or control nonimmune rabbit IgG followed by phycoerythrin-conjugated goat anti-rabbit secondary antibody (Jackson ImmunoResearch).

    Techniques: Activity Assay

    NEU3 catalytic activity in airway ECs. A–C : A549 cells were infected with increasing multiplicities of infection (MOIs) of Ad-NEU3-HA and cultured for 48 h. A : cells were lysed and the lysates were processed for immunoblotting with anti-hemagglutinin

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Human airway epithelia express catalytically active NEU3 sialidase

    doi: 10.1152/ajplung.00322.2013

    Figure Lengend Snippet: NEU3 catalytic activity in airway ECs. A–C : A549 cells were infected with increasing multiplicities of infection (MOIs) of Ad-NEU3-HA and cultured for 48 h. A : cells were lysed and the lysates were processed for immunoblotting with anti-hemagglutinin

    Article Snippet: Nonpermeabilized unstimulated 1HAEo− , CFTE29o− , 16HBE14o− , and BEAS-2B cells, SAECs, and A549 cells were stained with rabbit polyclonal anti-human NEU3 antibody (Novus Biologicals, Littleton, CO) or control nonimmune rabbit IgG followed by phycoerythrin-conjugated goat anti-rabbit secondary antibody (Jackson ImmunoResearch).

    Techniques: Activity Assay, Infection, Cell Culture

    Subcellular localization of NEU3. A : subconfluent A549 cells were probed with anti-NEU3 antibody ( I and ii ), or nonimmune IgG as a negative control ( iii ), and counterstained with DAPI (overlays in i and iii ). Arrows indicate the nuclear regions in NEU3-stained

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Human airway epithelia express catalytically active NEU3 sialidase

    doi: 10.1152/ajplung.00322.2013

    Figure Lengend Snippet: Subcellular localization of NEU3. A : subconfluent A549 cells were probed with anti-NEU3 antibody ( I and ii ), or nonimmune IgG as a negative control ( iii ), and counterstained with DAPI (overlays in i and iii ). Arrows indicate the nuclear regions in NEU3-stained

    Article Snippet: Nonpermeabilized unstimulated 1HAEo− , CFTE29o− , 16HBE14o− , and BEAS-2B cells, SAECs, and A549 cells were stained with rabbit polyclonal anti-human NEU3 antibody (Novus Biologicals, Littleton, CO) or control nonimmune rabbit IgG followed by phycoerythrin-conjugated goat anti-rabbit secondary antibody (Jackson ImmunoResearch).

    Techniques: Negative Control, Staining

    Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts

    Journal: Journal of Virology

    Article Title: Adenovirus Type 5 Early Region 1B 55K Oncoprotein-Dependent Degradation of Cellular Factor Daxx Is Required for Efficient Transformation of Primary Rodent Cells ▿

    doi: 10.1128/JVI.00440-11

    Figure Lengend Snippet: Construction and phenotypic analysis of the E1B-55K virus mutants H5 pm 4197 and H5 pm 4198. (A) A549 cells were infected with wild-type H5 pg 4100 or E1B-mutant viruses H5 pm 4149, H5 pm 4197, and H5 pm 4198 at a multiplicity of 50 FFU per cell, and whole-cell extracts

    Article Snippet: Primary BRK cells and BRK-derived cell lines (BRK1, AB120, and ABS1) , HEK293 cells , A549 cells (DSMZ ACC107; Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany), and the p53-negative human cell line H1299 ( , ) were grown in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS), 100 U of penicillin, and 100 μg of streptomycin per ml in a 5% CO2 atmosphere at 37°C.

    Techniques: Infection, Mutagenesis

    Confocal microscopic images of A549 cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.

    Journal: ACS Omega

    Article Title: Gemini Amphiphile-Based Lipoplexes for Efficient Gene Delivery: Synthesis, Formulation Development, Characterization, Gene Transfection, and Biodistribution Studies

    doi: 10.1021/acsomega.8b01014

    Figure Lengend Snippet: Confocal microscopic images of A549 cells treated with the lipolex formed from formulation 5d(chol) : first column shows the control group (cells without any lipoplex treatment), second column after 10 min, third column after 20 min, and fourth column after 30 min of lipoplex treatment of the cells. The bottom row images represent phase contrast images of the cells, whereas the upper ones are merged images showing blue-stained nucleus and the green dots surrounding the cytoplasm and entering the nucleus, at different time intervals.

    Article Snippet: HeLa and A549 cell lines were obtained from the National Centre for Cell Sciences (NCCS), Pune, India.

    Techniques: Staining

    FACS studies of the optimized formulations of 5b and 5d without cholesterol and with cholesterol [ 5b(chol) and 5d(chol) ] using GFP expression in A549 cells without FBS.

    Journal: ACS Omega

    Article Title: Gemini Amphiphile-Based Lipoplexes for Efficient Gene Delivery: Synthesis, Formulation Development, Characterization, Gene Transfection, and Biodistribution Studies

    doi: 10.1021/acsomega.8b01014

    Figure Lengend Snippet: FACS studies of the optimized formulations of 5b and 5d without cholesterol and with cholesterol [ 5b(chol) and 5d(chol) ] using GFP expression in A549 cells without FBS.

    Article Snippet: HeLa and A549 cell lines were obtained from the National Centre for Cell Sciences (NCCS), Pune, India.

    Techniques: FACS, Expressing

    Compiled results of percent cell viability of the optimized GA formulations (showing the best N/P results in transfection studies) in A549 cells, of the GAs showing the highest transfection efficacies.

    Journal: ACS Omega

    Article Title: Gemini Amphiphile-Based Lipoplexes for Efficient Gene Delivery: Synthesis, Formulation Development, Characterization, Gene Transfection, and Biodistribution Studies

    doi: 10.1021/acsomega.8b01014

    Figure Lengend Snippet: Compiled results of percent cell viability of the optimized GA formulations (showing the best N/P results in transfection studies) in A549 cells, of the GAs showing the highest transfection efficacies.

    Article Snippet: HeLa and A549 cell lines were obtained from the National Centre for Cell Sciences (NCCS), Pune, India.

    Techniques: Transfection

    Differential responses to carboplatin treatment in PC9 and A549 cells. A: Carboplatin-induced growth inhibition in PC9 and A549 cells using a sulforhodamine B assay 72 h post treatment. B: Western blot analysis of the levels of uncleaved PARP, cleaved PARP and cleaved (active) caspase 3 72 h post carboplatin treatment (0–200 µM) in PC9 and A549 cells. Actin was used as a loading control. C, D: Flow cytometric analysis of PC9 (C) and A549 cells (D) treated with carboplatin (100 µM) or vehicle. Apoptotic cells were identified by Annexin V-Alexafluor488 (λ Ex/Em = 495/519 nm) and necrotic cells by 7-AAD (λ Ex/Em = 546/647 nm). Population Q4 represents viable cells, whereas population Q3 represents apoptotic cells that have low 7-AAD fluorescence and stain with Annexin V. Population Q2 represents secondary apoptotic/necrotic cells.

    Journal: PLoS ONE

    Article Title: Preclinical Assessment of Carboplatin Treatment Efficacy in Lung Cancer by 18F-ICMT-11-Positron Emission Tomography

    doi: 10.1371/journal.pone.0091694

    Figure Lengend Snippet: Differential responses to carboplatin treatment in PC9 and A549 cells. A: Carboplatin-induced growth inhibition in PC9 and A549 cells using a sulforhodamine B assay 72 h post treatment. B: Western blot analysis of the levels of uncleaved PARP, cleaved PARP and cleaved (active) caspase 3 72 h post carboplatin treatment (0–200 µM) in PC9 and A549 cells. Actin was used as a loading control. C, D: Flow cytometric analysis of PC9 (C) and A549 cells (D) treated with carboplatin (100 µM) or vehicle. Apoptotic cells were identified by Annexin V-Alexafluor488 (λ Ex/Em = 495/519 nm) and necrotic cells by 7-AAD (λ Ex/Em = 546/647 nm). Population Q4 represents viable cells, whereas population Q3 represents apoptotic cells that have low 7-AAD fluorescence and stain with Annexin V. Population Q2 represents secondary apoptotic/necrotic cells.

    Article Snippet: In A549 cells, cell death was induced via the necrotic pathway, concurrent with minimal 18 F-ICMT-11 uptake, indicating great specificity of 18 F-ICMT-11 to trace apoptotic, but not necrotic mechanisms of cell death.

    Techniques: Inhibition, Sulforhodamine B Assay, Western Blot, Flow Cytometry, Fluorescence, Staining

    Temporal changes in cell death markers and 18 F-ICMT-11 uptake after carboplatin treatment. A: Time course of changes in caspase 3/7 activity following carboplatin treatment. B: Western blot analysis of the levels of uncleaved PARP, cleaved PARP and cleaved (active) caspase 3 post 50 µM carboplatin treatment (0–96 h) in PC9 (i) and A549 cells (ii). C: Temporal changes in 18 F-ICMT-11 uptake in cells following carboplatin treatment. D: Correlation between caspase 3 activity and 18 F-ICMT-11 uptake in PC9 cells.

    Journal: PLoS ONE

    Article Title: Preclinical Assessment of Carboplatin Treatment Efficacy in Lung Cancer by 18F-ICMT-11-Positron Emission Tomography

    doi: 10.1371/journal.pone.0091694

    Figure Lengend Snippet: Temporal changes in cell death markers and 18 F-ICMT-11 uptake after carboplatin treatment. A: Time course of changes in caspase 3/7 activity following carboplatin treatment. B: Western blot analysis of the levels of uncleaved PARP, cleaved PARP and cleaved (active) caspase 3 post 50 µM carboplatin treatment (0–96 h) in PC9 (i) and A549 cells (ii). C: Temporal changes in 18 F-ICMT-11 uptake in cells following carboplatin treatment. D: Correlation between caspase 3 activity and 18 F-ICMT-11 uptake in PC9 cells.

    Article Snippet: In A549 cells, cell death was induced via the necrotic pathway, concurrent with minimal 18 F-ICMT-11 uptake, indicating great specificity of 18 F-ICMT-11 to trace apoptotic, but not necrotic mechanisms of cell death.

    Techniques: Activity Assay, Western Blot

    Voxel-wise analysis of 18 F-ICMT-11 PET imaging data by PVIS. The intensities of all voxels within the tumour ROIs were computed and expressed as histogram plots of normalized voxel intensity versus the number of voxels. A, B: Typical data from three representative animals (vehicle, 24 h or 48 h carboplatin-treated) for PC9 (A) and A549 (B) are shown. C, D: The statistical comparison of 95 th percentile voxel intensities for PC9 (C) and A549 (D) was performed using Prism v5.0 software (GraphPad). Mean ± SD ( n = 4–6 animals per group).*, P

    Journal: PLoS ONE

    Article Title: Preclinical Assessment of Carboplatin Treatment Efficacy in Lung Cancer by 18F-ICMT-11-Positron Emission Tomography

    doi: 10.1371/journal.pone.0091694

    Figure Lengend Snippet: Voxel-wise analysis of 18 F-ICMT-11 PET imaging data by PVIS. The intensities of all voxels within the tumour ROIs were computed and expressed as histogram plots of normalized voxel intensity versus the number of voxels. A, B: Typical data from three representative animals (vehicle, 24 h or 48 h carboplatin-treated) for PC9 (A) and A549 (B) are shown. C, D: The statistical comparison of 95 th percentile voxel intensities for PC9 (C) and A549 (D) was performed using Prism v5.0 software (GraphPad). Mean ± SD ( n = 4–6 animals per group).*, P

    Article Snippet: In A549 cells, cell death was induced via the necrotic pathway, concurrent with minimal 18 F-ICMT-11 uptake, indicating great specificity of 18 F-ICMT-11 to trace apoptotic, but not necrotic mechanisms of cell death.

    Techniques: Positron Emission Tomography, Imaging, Software

    18 F-ICMT-11 PET image analysis of PC9 and A549 xenografts in vehicle and carboplatin-treated mice. A, B: Tumour volumes recorded by calliper measurements of PC9 (A) and A549 tumours (B) pre- and post-carboplatin treatment as indicated. Data shown are mean ± SD of % volume compared to baseline ( n = 4). *, P

    Journal: PLoS ONE

    Article Title: Preclinical Assessment of Carboplatin Treatment Efficacy in Lung Cancer by 18F-ICMT-11-Positron Emission Tomography

    doi: 10.1371/journal.pone.0091694

    Figure Lengend Snippet: 18 F-ICMT-11 PET image analysis of PC9 and A549 xenografts in vehicle and carboplatin-treated mice. A, B: Tumour volumes recorded by calliper measurements of PC9 (A) and A549 tumours (B) pre- and post-carboplatin treatment as indicated. Data shown are mean ± SD of % volume compared to baseline ( n = 4). *, P

    Article Snippet: In A549 cells, cell death was induced via the necrotic pathway, concurrent with minimal 18 F-ICMT-11 uptake, indicating great specificity of 18 F-ICMT-11 to trace apoptotic, but not necrotic mechanisms of cell death.

    Techniques: Positron Emission Tomography, Mouse Assay

    ( A ) Antitumor effect of CK and CK mixed micelles to A549 cell on different concentrations. * P

    Journal: International Journal of Nanomedicine

    Article Title: Ascorbyl palmitate/d-α-tocopheryl polyethylene glycol 1000 succinate monoester mixed micelles for prolonged circulation and targeted delivery of compound K for antilung cancer therapy in vitro and in vivo

    doi: 10.2147/IJN.S119226

    Figure Lengend Snippet: ( A ) Antitumor effect of CK and CK mixed micelles to A549 cell on different concentrations. * P

    Article Snippet: Briefly, after being treated with CK or CK mixed micelles, A549 cells were permeabilized by Tris-HCl and ethylenediaminetetraacetic acid (EDTA) for 10 min.

    Techniques:

    Fluorescence microscopy of A549 cell following 4 h of treatment with C-6 or C-6 mixed micelles (n=3). Abbreviations: C-6, coumarin-6; DAPI, 4′,6-diamidino-2-phenylindole; h, hours.

    Journal: International Journal of Nanomedicine

    Article Title: Ascorbyl palmitate/d-α-tocopheryl polyethylene glycol 1000 succinate monoester mixed micelles for prolonged circulation and targeted delivery of compound K for antilung cancer therapy in vitro and in vivo

    doi: 10.2147/IJN.S119226

    Figure Lengend Snippet: Fluorescence microscopy of A549 cell following 4 h of treatment with C-6 or C-6 mixed micelles (n=3). Abbreviations: C-6, coumarin-6; DAPI, 4′,6-diamidino-2-phenylindole; h, hours.

    Article Snippet: Briefly, after being treated with CK or CK mixed micelles, A549 cells were permeabilized by Tris-HCl and ethylenediaminetetraacetic acid (EDTA) for 10 min.

    Techniques: Fluorescence, Microscopy

    Confocal microscopy of A549 cell following 4 h of treatment with C-6 or C-6 mixed micelles (n=3). Abbreviations: C-6, coumarin-6; DAPI, 4′,6-diamidino-2-phenylindole; h, hours.

    Journal: International Journal of Nanomedicine

    Article Title: Ascorbyl palmitate/d-α-tocopheryl polyethylene glycol 1000 succinate monoester mixed micelles for prolonged circulation and targeted delivery of compound K for antilung cancer therapy in vitro and in vivo

    doi: 10.2147/IJN.S119226

    Figure Lengend Snippet: Confocal microscopy of A549 cell following 4 h of treatment with C-6 or C-6 mixed micelles (n=3). Abbreviations: C-6, coumarin-6; DAPI, 4′,6-diamidino-2-phenylindole; h, hours.

    Article Snippet: Briefly, after being treated with CK or CK mixed micelles, A549 cells were permeabilized by Tris-HCl and ethylenediaminetetraacetic acid (EDTA) for 10 min.

    Techniques: Confocal Microscopy

    Cell cycle analysis assay (green G1, brown S, and blue G2 phase). ( A ) and wound healing assay ( B ) on A549 cell. Notes: n=3, mean ± SE. Scale 50 μm. Abbreviations: CK, compound K; SE, standard error.

    Journal: International Journal of Nanomedicine

    Article Title: Ascorbyl palmitate/d-α-tocopheryl polyethylene glycol 1000 succinate monoester mixed micelles for prolonged circulation and targeted delivery of compound K for antilung cancer therapy in vitro and in vivo

    doi: 10.2147/IJN.S119226

    Figure Lengend Snippet: Cell cycle analysis assay (green G1, brown S, and blue G2 phase). ( A ) and wound healing assay ( B ) on A549 cell. Notes: n=3, mean ± SE. Scale 50 μm. Abbreviations: CK, compound K; SE, standard error.

    Article Snippet: Briefly, after being treated with CK or CK mixed micelles, A549 cells were permeabilized by Tris-HCl and ethylenediaminetetraacetic acid (EDTA) for 10 min.

    Techniques: Cell Cycle Assay, Wound Healing Assay

    Combinatorial CRISPR screens reveal metabolic network dependencies (A) SKO fitness scores for HeLa cells, plotted as f g (day −1 ), with a more negative score representing a loss in fitness with SKO. Plotted as mean ± SD. (B) Multi-isoform family member fitness scores and gene expression for HeLa (top) and A549 (bottom) cells. (C) Relative comparison of SKO fitness scores (f g ) across both cells. (D) Relative comparison of genetic interaction scores (π gg .

    Journal: Molecular cell

    Article Title: Combinatorial CRISPR-Cas9 metabolic screens reveal critical redox control points dependent on the KEAP1-NRF2 regulatory axis

    doi: 10.1016/j.molcel.2018.01.017

    Figure Lengend Snippet: Combinatorial CRISPR screens reveal metabolic network dependencies (A) SKO fitness scores for HeLa cells, plotted as f g (day −1 ), with a more negative score representing a loss in fitness with SKO. Plotted as mean ± SD. (B) Multi-isoform family member fitness scores and gene expression for HeLa (top) and A549 (bottom) cells. (C) Relative comparison of SKO fitness scores (f g ) across both cells. (D) Relative comparison of genetic interaction scores (π gg .

    Article Snippet: CRISPR Cas9 nuclease stable expressing HeLa and A549 cells were obtained from GeneCopoeia and grown in DMEM medium with 10% FBS and Antibiotic-Antimycotic.

    Techniques: CRISPR, Expressing

    Cell cycle distribution of A549 cells line and A549 tumor spheres on effects of chemotherapeutic drugs

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells

    doi:

    Figure Lengend Snippet: Cell cycle distribution of A549 cells line and A549 tumor spheres on effects of chemotherapeutic drugs

    Article Snippet: Protein extracts of A549 cells line, A549 secondary spheres and HBE were resolved by 12% SDS-PAGE and transferred on PVDF (Millipore, USA) membranes using ECL Semi-dry Blotters (TE70PWR, USA) electro transfer system.

    Techniques:

    Isolation and identification of LCSCs. A. Three types of colonies formed by A549 cells. (400×). B. Immunostaining of stem cell markers in secondary spheres. Nuclei were stained using DAPI (blue). LCM, scale bar = 25 10, 50, 75, 10, 30 μm

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells

    doi:

    Figure Lengend Snippet: Isolation and identification of LCSCs. A. Three types of colonies formed by A549 cells. (400×). B. Immunostaining of stem cell markers in secondary spheres. Nuclei were stained using DAPI (blue). LCM, scale bar = 25 10, 50, 75, 10, 30 μm

    Article Snippet: Protein extracts of A549 cells line, A549 secondary spheres and HBE were resolved by 12% SDS-PAGE and transferred on PVDF (Millipore, USA) membranes using ECL Semi-dry Blotters (TE70PWR, USA) electro transfer system.

    Techniques: Isolation, Immunostaining, Staining, Laser Capture Microdissection

    Enhanced expression of GST-π, MRP1 and LRP in A549 spheres. A. Western-blot of GST-π, MRP1 and LRP in HEB, A549 cells and A549 spheres. B. Immunofluorescence of GST-π expression in HEB, A549 cells and A549 spheres using confocal

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells

    doi:

    Figure Lengend Snippet: Enhanced expression of GST-π, MRP1 and LRP in A549 spheres. A. Western-blot of GST-π, MRP1 and LRP in HEB, A549 cells and A549 spheres. B. Immunofluorescence of GST-π expression in HEB, A549 cells and A549 spheres using confocal

    Article Snippet: Protein extracts of A549 cells line, A549 secondary spheres and HBE were resolved by 12% SDS-PAGE and transferred on PVDF (Millipore, USA) membranes using ECL Semi-dry Blotters (TE70PWR, USA) electro transfer system.

    Techniques: Expressing, Western Blot, Immunofluorescence

    Effect of the IC50 concentration of Cisplatin (DDP) and Gemcitabine (GEM) on the cell cycle profile of A549 cells line and A549 spheres. A. A549 cells line; B. A549 cells line/DDP; C. A549 cells line/GEM; D. A549 cancer spheres; E. A549 cancer spheres/DDP;

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells

    doi:

    Figure Lengend Snippet: Effect of the IC50 concentration of Cisplatin (DDP) and Gemcitabine (GEM) on the cell cycle profile of A549 cells line and A549 spheres. A. A549 cells line; B. A549 cells line/DDP; C. A549 cells line/GEM; D. A549 cancer spheres; E. A549 cancer spheres/DDP;

    Article Snippet: Protein extracts of A549 cells line, A549 secondary spheres and HBE were resolved by 12% SDS-PAGE and transferred on PVDF (Millipore, USA) membranes using ECL Semi-dry Blotters (TE70PWR, USA) electro transfer system.

    Techniques: Concentration Assay

    Measurement of the sensitivity of A549 cancer spheres and A549 cells line to anticancer drugs (DDP and GEM). A. The lethal doses (IC50) of DDP (left) and GEM (right) in A549 cancer spheres and A549 cells line, The data shown represent the mean ±

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells

    doi:

    Figure Lengend Snippet: Measurement of the sensitivity of A549 cancer spheres and A549 cells line to anticancer drugs (DDP and GEM). A. The lethal doses (IC50) of DDP (left) and GEM (right) in A549 cancer spheres and A549 cells line, The data shown represent the mean ±

    Article Snippet: Protein extracts of A549 cells line, A549 secondary spheres and HBE were resolved by 12% SDS-PAGE and transferred on PVDF (Millipore, USA) membranes using ECL Semi-dry Blotters (TE70PWR, USA) electro transfer system.

    Techniques:

    A549 cancer spheres have high express CD133 and more self renewal capability. A. The morphology of tumor spheres. a. Primary tumor spheres formed in the CSM medium. b. Secondary spheres derived from single parental cells of primary tumor spheres under

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Enhanced expression of stem cell markers and drug resistance in sphere-forming non-small cell lung cancer cells

    doi:

    Figure Lengend Snippet: A549 cancer spheres have high express CD133 and more self renewal capability. A. The morphology of tumor spheres. a. Primary tumor spheres formed in the CSM medium. b. Secondary spheres derived from single parental cells of primary tumor spheres under

    Article Snippet: Protein extracts of A549 cells line, A549 secondary spheres and HBE were resolved by 12% SDS-PAGE and transferred on PVDF (Millipore, USA) membranes using ECL Semi-dry Blotters (TE70PWR, USA) electro transfer system.

    Techniques: Derivative Assay

    The dual addition of DL-PDMP and D-NMAPPD to A549 cell culture induced the possibility of Cer channel formation via endogenous C16:0-Cer accumulation in the lysosome membrane as an initial step prior to initiation of necrotic cell death. The findings described in this study are summarized with the findings of Ref. [7] . As for the morphology, the dual addition of DL-PDMP/D-NMAPPD led to black circular structures of 10–20 nm, interpreted as stain-filled cylindrical channels on transmission electron microscopy of the lysosome-rich fraction.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

    doi: 10.1016/j.bbrep.2017.02.010

    Figure Lengend Snippet: The dual addition of DL-PDMP and D-NMAPPD to A549 cell culture induced the possibility of Cer channel formation via endogenous C16:0-Cer accumulation in the lysosome membrane as an initial step prior to initiation of necrotic cell death. The findings described in this study are summarized with the findings of Ref. [7] . As for the morphology, the dual addition of DL-PDMP/D-NMAPPD led to black circular structures of 10–20 nm, interpreted as stain-filled cylindrical channels on transmission electron microscopy of the lysosome-rich fraction.

    Article Snippet: 2.2 A549 cell culture, induction of Cer accumulation A549 cells (human lung adenocarcinoma cell line) were grown in humidified air with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (including 8.5 µM free fatty acids) prepared from Sigma D5796 including 400 µM L-serine, containing fetal bovine serum (FBS) at a concentration of 10% (v/v), at 37 °C.

    Techniques: Cell Culture, Staining, Transmission Assay, Electron Microscopy

    The individual addition of NAC or CA-074Me to the dual addition of DL-PDMP and D-NMAPPD did not significatly modify C16:0-Cer/sphinganine/sphingosine contents compared with the dual addition. The induction of Cer accumulation and extraction/analysis of C16:0-Cer/sphinganine/sphingosine in the homogenate of A549 cells were achieved as described in Methods. A – C , levels of C16:0-Cer/sphinganine/sphingosine in A549 cells 5 h after the control or dual/triple addition. Data are presented as the mean values±S.D. of three independent experiments; **P

    Journal: Biochemistry and Biophysics Reports

    Article Title: Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

    doi: 10.1016/j.bbrep.2017.02.010

    Figure Lengend Snippet: The individual addition of NAC or CA-074Me to the dual addition of DL-PDMP and D-NMAPPD did not significatly modify C16:0-Cer/sphinganine/sphingosine contents compared with the dual addition. The induction of Cer accumulation and extraction/analysis of C16:0-Cer/sphinganine/sphingosine in the homogenate of A549 cells were achieved as described in Methods. A – C , levels of C16:0-Cer/sphinganine/sphingosine in A549 cells 5 h after the control or dual/triple addition. Data are presented as the mean values±S.D. of three independent experiments; **P

    Article Snippet: 2.2 A549 cell culture, induction of Cer accumulation A549 cells (human lung adenocarcinoma cell line) were grown in humidified air with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (including 8.5 µM free fatty acids) prepared from Sigma D5796 including 400 µM L-serine, containing fetal bovine serum (FBS) at a concentration of 10% (v/v), at 37 °C.

    Techniques:

    The analysis of LAMP-2, Bax, and β-actin in the homogenate or lysosome-rich fraction from A549 cells by Western blotting. Western blotting analysis was achieved as described in Methods. LAMP-2/β-actin was high in the lysosome-rich fraction and LAMP-2 protein was concentrated in the lysosome-rich fraction. However, Bax protein was not detected in the lysosome-rich fraction. Data are presented as the mean values±S.D. of three independent experiments;*P

    Journal: Biochemistry and Biophysics Reports

    Article Title: Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

    doi: 10.1016/j.bbrep.2017.02.010

    Figure Lengend Snippet: The analysis of LAMP-2, Bax, and β-actin in the homogenate or lysosome-rich fraction from A549 cells by Western blotting. Western blotting analysis was achieved as described in Methods. LAMP-2/β-actin was high in the lysosome-rich fraction and LAMP-2 protein was concentrated in the lysosome-rich fraction. However, Bax protein was not detected in the lysosome-rich fraction. Data are presented as the mean values±S.D. of three independent experiments;*P

    Article Snippet: 2.2 A549 cell culture, induction of Cer accumulation A549 cells (human lung adenocarcinoma cell line) were grown in humidified air with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (including 8.5 µM free fatty acids) prepared from Sigma D5796 including 400 µM L-serine, containing fetal bovine serum (FBS) at a concentration of 10% (v/v), at 37 °C.

    Techniques: Western Blot

    The dual addition of DL-PDMP and D-NMAPPD caused an additional increase in C16:0-Cer, dihydro-C16:0-Cer or cholesterol contents in the lysosome-rich fraction of A549 cells 2 h afterwards, compared with the control or individual addition. The induction of Cer accumulation and extraction/analysis of Cers/dihydro-Cer/cholesterol in the homogenate or the lysosome-rich fraction were achieved as described in Methods. A - D , levels of Cer and dihydro-Cer species in the homogenate or the lysosomal rich fraction of A549 cells 2 h after the individual or dual addition. E, levels of cholesterol in the homogenate or the lysosome-rich fraction of A549 cells 2 h after the individual or dual addition. Data are presented as the mean values±S.D. of three independent experiments; *P

    Journal: Biochemistry and Biophysics Reports

    Article Title: Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

    doi: 10.1016/j.bbrep.2017.02.010

    Figure Lengend Snippet: The dual addition of DL-PDMP and D-NMAPPD caused an additional increase in C16:0-Cer, dihydro-C16:0-Cer or cholesterol contents in the lysosome-rich fraction of A549 cells 2 h afterwards, compared with the control or individual addition. The induction of Cer accumulation and extraction/analysis of Cers/dihydro-Cer/cholesterol in the homogenate or the lysosome-rich fraction were achieved as described in Methods. A - D , levels of Cer and dihydro-Cer species in the homogenate or the lysosomal rich fraction of A549 cells 2 h after the individual or dual addition. E, levels of cholesterol in the homogenate or the lysosome-rich fraction of A549 cells 2 h after the individual or dual addition. Data are presented as the mean values±S.D. of three independent experiments; *P

    Article Snippet: 2.2 A549 cell culture, induction of Cer accumulation A549 cells (human lung adenocarcinoma cell line) were grown in humidified air with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (including 8.5 µM free fatty acids) prepared from Sigma D5796 including 400 µM L-serine, containing fetal bovine serum (FBS) at a concentration of 10% (v/v), at 37 °C.

    Techniques:

    Confocal microscopy images of incubated with poly-G-quadruplexes-ThT probe treated A549 ( a ) and HepG2 ( b ). Scale bar: 20 µm. ( c ) The normalized fluorescence intensity of individual cells was quantified from ( a ) and ( b ). Error bars indicate SD, n = 5. **P

    Journal: Scientific Reports

    Article Title: Simultaneous Monitoring of Cell-surface Receptor and Tumor-targeted Photodynamic Therapy via TdT-initiated Poly-G-Quadruplexes

    doi: 10.1038/s41598-018-23902-5

    Figure Lengend Snippet: Confocal microscopy images of incubated with poly-G-quadruplexes-ThT probe treated A549 ( a ) and HepG2 ( b ). Scale bar: 20 µm. ( c ) The normalized fluorescence intensity of individual cells was quantified from ( a ) and ( b ). Error bars indicate SD, n = 5. **P

    Article Snippet: Cell culture A549 cells (human lung adenocarcinoma epithelial cell line) and HepG2 cells (human hepatocellular carcinoma cell line) were cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO) with 10% fetal bovine serum (FBS) and 0.5 mg/mL penicillin-streptomycin in a humidified incubator at 37 °C under a 5% CO2 atmosphere.

    Techniques: Confocal Microscopy, Incubation, Fluorescence

    ( a ) Fluorescence emission spectra of ThT in the presence of poly-G-quadruplexes or mono-G-quadruplex. ( b ) Time-dependent photo-bleaching effects of the fluorescent intensity of the poly-G-quadruplexes-ThT complex or FITC, fluorescence intensity was normalized to the maximum intensity of each fluorophore and then plotted against exposure time. Confocal images of A549 cells treated with poly-G-quadruplexes ( c ) and mono-G-quadruplex ( d ). Scale bar: 20 µm. ( e ) The normalized fluorescence intensity of individual cells was quantified from ( c ) and ( d ). Error bars indicate SD, n = 5. **P

    Journal: Scientific Reports

    Article Title: Simultaneous Monitoring of Cell-surface Receptor and Tumor-targeted Photodynamic Therapy via TdT-initiated Poly-G-Quadruplexes

    doi: 10.1038/s41598-018-23902-5

    Figure Lengend Snippet: ( a ) Fluorescence emission spectra of ThT in the presence of poly-G-quadruplexes or mono-G-quadruplex. ( b ) Time-dependent photo-bleaching effects of the fluorescent intensity of the poly-G-quadruplexes-ThT complex or FITC, fluorescence intensity was normalized to the maximum intensity of each fluorophore and then plotted against exposure time. Confocal images of A549 cells treated with poly-G-quadruplexes ( c ) and mono-G-quadruplex ( d ). Scale bar: 20 µm. ( e ) The normalized fluorescence intensity of individual cells was quantified from ( c ) and ( d ). Error bars indicate SD, n = 5. **P

    Article Snippet: Cell culture A549 cells (human lung adenocarcinoma epithelial cell line) and HepG2 cells (human hepatocellular carcinoma cell line) were cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO) with 10% fetal bovine serum (FBS) and 0.5 mg/mL penicillin-streptomycin in a humidified incubator at 37 °C under a 5% CO2 atmosphere.

    Techniques: Fluorescence

    ( a ) Schematic illustration of TMPyP4 competes with ThT in combination with poly-G-quadruplexes on cell-surface. ( b ) Confocal imaging of poly-G-quadruplexes-ThT treated A549 cells with increasing concentrations of TMPyP4 (0, 5, 10, 20, 50 µM). Scale bar: 20 µm. ( c ) The normalized fluorescence intensity of individual cells was quantified from ( b ). Error bars indicate SD, n = 5. ( d ) Cell-surface normalized fluorescence intensity of the ThT at 485nm under increasing concentrations of TMPyP4 (0, 5, 10, 20, 50 µM) by using microplate reader.

    Journal: Scientific Reports

    Article Title: Simultaneous Monitoring of Cell-surface Receptor and Tumor-targeted Photodynamic Therapy via TdT-initiated Poly-G-Quadruplexes

    doi: 10.1038/s41598-018-23902-5

    Figure Lengend Snippet: ( a ) Schematic illustration of TMPyP4 competes with ThT in combination with poly-G-quadruplexes on cell-surface. ( b ) Confocal imaging of poly-G-quadruplexes-ThT treated A549 cells with increasing concentrations of TMPyP4 (0, 5, 10, 20, 50 µM). Scale bar: 20 µm. ( c ) The normalized fluorescence intensity of individual cells was quantified from ( b ). Error bars indicate SD, n = 5. ( d ) Cell-surface normalized fluorescence intensity of the ThT at 485nm under increasing concentrations of TMPyP4 (0, 5, 10, 20, 50 µM) by using microplate reader.

    Article Snippet: Cell culture A549 cells (human lung adenocarcinoma epithelial cell line) and HepG2 cells (human hepatocellular carcinoma cell line) were cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO) with 10% fetal bovine serum (FBS) and 0.5 mg/mL penicillin-streptomycin in a humidified incubator at 37 °C under a 5% CO2 atmosphere.

    Techniques: Imaging, Fluorescence

    ( a ) Time-dependent ROS generation by poly-G-quadruplexes-TMPyP4 under light irradiation was measured using DCF fluorescence. ( b ) Schematic illustration of the ROS generation from the poly-G-quadruplexes-TMPyP4 system under light irradiation. ( c ) The fluorescence microscopy images of light-induced ROS production of A549 cells treated with poly-G-quadruplexes-ThT-TMPyP4 with or without light irradiation, scale bar: 20 µm. ( d ) Characterization of the selective cytotoxicity of TMPyP4 delivered by the poly-G-quadruplexes conjugated probe to A549 cells (red), HepG2 (blue). Error bars indicate SD, n = 3. **P

    Journal: Scientific Reports

    Article Title: Simultaneous Monitoring of Cell-surface Receptor and Tumor-targeted Photodynamic Therapy via TdT-initiated Poly-G-Quadruplexes

    doi: 10.1038/s41598-018-23902-5

    Figure Lengend Snippet: ( a ) Time-dependent ROS generation by poly-G-quadruplexes-TMPyP4 under light irradiation was measured using DCF fluorescence. ( b ) Schematic illustration of the ROS generation from the poly-G-quadruplexes-TMPyP4 system under light irradiation. ( c ) The fluorescence microscopy images of light-induced ROS production of A549 cells treated with poly-G-quadruplexes-ThT-TMPyP4 with or without light irradiation, scale bar: 20 µm. ( d ) Characterization of the selective cytotoxicity of TMPyP4 delivered by the poly-G-quadruplexes conjugated probe to A549 cells (red), HepG2 (blue). Error bars indicate SD, n = 3. **P

    Article Snippet: Cell culture A549 cells (human lung adenocarcinoma epithelial cell line) and HepG2 cells (human hepatocellular carcinoma cell line) were cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich, St. Louis, MO) with 10% fetal bovine serum (FBS) and 0.5 mg/mL penicillin-streptomycin in a humidified incubator at 37 °C under a 5% CO2 atmosphere.

    Techniques: Irradiation, Fluorescence, Microscopy

    Suppressive effect of miR-143 on the proliferation of non-small cell lunger cancer cells. (A) Successful transfection of A549 cells with miR-143 was confirmed by quantitative polymerase chain reaction. (B) The proliferation of A549 cells transfected with

    Journal: Experimental and Therapeutic Medicine

    Article Title: miR-143 suppresses the proliferation of NSCLC cells by inhibiting the epidermal growth factor receptor

    doi: 10.3892/etm.2016.3555

    Figure Lengend Snippet: Suppressive effect of miR-143 on the proliferation of non-small cell lunger cancer cells. (A) Successful transfection of A549 cells with miR-143 was confirmed by quantitative polymerase chain reaction. (B) The proliferation of A549 cells transfected with

    Article Snippet: A549 cell proliferation was assessed using a BrdU Cell Proliferation ELISA kit (EMD Millipore, Billerica, MA, USA).

    Techniques: Transfection, Real-time Polymerase Chain Reaction

    Similar effects were observed for EGFR inhibition and microRNA-143 overexpression. The (A) mRNA and (B and C) protein expression levels of EGFR were significantly reduced in the A549 cells transfected with EGFR shRNA compared with the cells transfected

    Journal: Experimental and Therapeutic Medicine

    Article Title: miR-143 suppresses the proliferation of NSCLC cells by inhibiting the epidermal growth factor receptor

    doi: 10.3892/etm.2016.3555

    Figure Lengend Snippet: Similar effects were observed for EGFR inhibition and microRNA-143 overexpression. The (A) mRNA and (B and C) protein expression levels of EGFR were significantly reduced in the A549 cells transfected with EGFR shRNA compared with the cells transfected

    Article Snippet: A549 cell proliferation was assessed using a BrdU Cell Proliferation ELISA kit (EMD Millipore, Billerica, MA, USA).

    Techniques: Inhibition, Over Expression, Expressing, Transfection, shRNA

    Suppressive effect of miR-143 on non-small cell lung cancer cell migration and invasion. The migration and invasion of A549 cells was assessed using a Transwell assay. The (A) migration and (B) invasion of A549 cells transfected with miR-143 were significantly

    Journal: Experimental and Therapeutic Medicine

    Article Title: miR-143 suppresses the proliferation of NSCLC cells by inhibiting the epidermal growth factor receptor

    doi: 10.3892/etm.2016.3555

    Figure Lengend Snippet: Suppressive effect of miR-143 on non-small cell lung cancer cell migration and invasion. The migration and invasion of A549 cells was assessed using a Transwell assay. The (A) migration and (B) invasion of A549 cells transfected with miR-143 were significantly

    Article Snippet: A549 cell proliferation was assessed using a BrdU Cell Proliferation ELISA kit (EMD Millipore, Billerica, MA, USA).

    Techniques: Migration, Transwell Assay, Transfection

    IFA staining of influenza virus A-infected cells from a clinical specimen. (A) Mv1Lu cells. (B) Mixed A549-Mv1Lu cells. (C) pRhMK cells. Magnification, ×100.

    Journal: Journal of Clinical Microbiology

    Article Title: Mink Lung Cells and Mixed Mink Lung and A549 Cells for Rapid Detection of Influenza Virus and Other Respiratory Viruses

    doi:

    Figure Lengend Snippet: IFA staining of influenza virus A-infected cells from a clinical specimen. (A) Mv1Lu cells. (B) Mixed A549-Mv1Lu cells. (C) pRhMK cells. Magnification, ×100.

    Article Snippet: It is very unlikely, but some strains of influenza virus A might grow extremely well in A549 cells.

    Techniques: Immunofluorescence, Staining, Infection

    MTT assays. A549 cells were treated with increasing concentration of XAV939 (0.1, 0.5, 1,5 and 10 µM) for 24, 4, 72 and 96 h. The result was shown as relative cell viability per concentration at each time point.

    Journal: Oncology Letters

    Article Title: XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway

    doi: 10.3892/ol.2018.8491

    Figure Lengend Snippet: MTT assays. A549 cells were treated with increasing concentration of XAV939 (0.1, 0.5, 1,5 and 10 µM) for 24, 4, 72 and 96 h. The result was shown as relative cell viability per concentration at each time point.

    Article Snippet: A549 cells treated with XAV939 or DDP exhibited decreased the expression of TNKS protein compared with untreated control cells. c-Myc, a downstream target of β-catenin, was also downregulated.

    Techniques: MTT Assay, Concentration Assay

    Localization of β-catenin immunofluorescence in response to treatment with NaCl (control), 0., 1 and 10 µmol/l XAV939 in A549 cells (magnification, ×400).

    Journal: Oncology Letters

    Article Title: XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway

    doi: 10.3892/ol.2018.8491

    Figure Lengend Snippet: Localization of β-catenin immunofluorescence in response to treatment with NaCl (control), 0., 1 and 10 µmol/l XAV939 in A549 cells (magnification, ×400).

    Article Snippet: A549 cells treated with XAV939 or DDP exhibited decreased the expression of TNKS protein compared with untreated control cells. c-Myc, a downstream target of β-catenin, was also downregulated.

    Techniques: Immunofluorescence

    c-Myc mRNA expression at different XAV939 concentrations as detected by RT-sqPCR. (A) RT-PCR gel (B) and relative expression of β-catenin mRNA in A549 cells, treated with different XAV939 concentrations. *P

    Journal: Oncology Letters

    Article Title: XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway

    doi: 10.3892/ol.2018.8491

    Figure Lengend Snippet: c-Myc mRNA expression at different XAV939 concentrations as detected by RT-sqPCR. (A) RT-PCR gel (B) and relative expression of β-catenin mRNA in A549 cells, treated with different XAV939 concentrations. *P

    Article Snippet: A549 cells treated with XAV939 or DDP exhibited decreased the expression of TNKS protein compared with untreated control cells. c-Myc, a downstream target of β-catenin, was also downregulated.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    Εffect of XAV939 on A549 cell migration was detected via wound-healing assay. The wound widths in the different XAV939 treatment groups (0.1, 0.5, 1, 5, 10 µmol/l) after 24 h were all significantly wider than those of the control group (magnification, ×200).

    Journal: Oncology Letters

    Article Title: XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway

    doi: 10.3892/ol.2018.8491

    Figure Lengend Snippet: Εffect of XAV939 on A549 cell migration was detected via wound-healing assay. The wound widths in the different XAV939 treatment groups (0.1, 0.5, 1, 5, 10 µmol/l) after 24 h were all significantly wider than those of the control group (magnification, ×200).

    Article Snippet: A549 cells treated with XAV939 or DDP exhibited decreased the expression of TNKS protein compared with untreated control cells. c-Myc, a downstream target of β-catenin, was also downregulated.

    Techniques: Migration, Wound Healing Assay

    β-catenin mRNA expression at different XAV939 concentrations as detected by RT-sqPCR. (A) RT-PCR gel (B) and relative expression of β-catenin mRNA in A549 cells, treated with different XAV939 concentrations. *P

    Journal: Oncology Letters

    Article Title: XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway

    doi: 10.3892/ol.2018.8491

    Figure Lengend Snippet: β-catenin mRNA expression at different XAV939 concentrations as detected by RT-sqPCR. (A) RT-PCR gel (B) and relative expression of β-catenin mRNA in A549 cells, treated with different XAV939 concentrations. *P

    Article Snippet: A549 cells treated with XAV939 or DDP exhibited decreased the expression of TNKS protein compared with untreated control cells. c-Myc, a downstream target of β-catenin, was also downregulated.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    (A) XAV939 inhibits the clonogenicity of A549 cells in vitro . (B) Treatment of the A549 cell line with 0.1, 1 and 10 µmol/l XAV939 significantly inhibited colony formation compared with the control. *P

    Journal: Oncology Letters

    Article Title: XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway

    doi: 10.3892/ol.2018.8491

    Figure Lengend Snippet: (A) XAV939 inhibits the clonogenicity of A549 cells in vitro . (B) Treatment of the A549 cell line with 0.1, 1 and 10 µmol/l XAV939 significantly inhibited colony formation compared with the control. *P

    Article Snippet: A549 cells treated with XAV939 or DDP exhibited decreased the expression of TNKS protein compared with untreated control cells. c-Myc, a downstream target of β-catenin, was also downregulated.

    Techniques: In Vitro

    Expression of TNKS in three lung adenocarcinoma cell lines. (A) Western blot analysis and (B) quantification of this analysis demonstrated that the level of TNKS protein expression in A549 cells was significantly higher compared with that in Calu-3 and SK-LU-1 cells. *P

    Journal: Oncology Letters

    Article Title: XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway

    doi: 10.3892/ol.2018.8491

    Figure Lengend Snippet: Expression of TNKS in three lung adenocarcinoma cell lines. (A) Western blot analysis and (B) quantification of this analysis demonstrated that the level of TNKS protein expression in A549 cells was significantly higher compared with that in Calu-3 and SK-LU-1 cells. *P

    Article Snippet: A549 cells treated with XAV939 or DDP exhibited decreased the expression of TNKS protein compared with untreated control cells. c-Myc, a downstream target of β-catenin, was also downregulated.

    Techniques: Expressing, Western Blot

    The effects of calcitriol on A549 cells. a Effect of calcitriol treatment on virus growth kinetic in A549 cells. Cell culture supernatants were harvested every 24 h from infected cells and the viral load was titrated using the TCID50 assay. Real-time PCR was used to measure the effect of calcitriol treatment on the mRNA expression levels of the viral M gene ( b ), IL-6 ( c ), and IFN-β ( d ) in A549 cells. β-actin was used as an internal control. The data are expressed as mean ± SEM of triplicate samples

    Journal: Virology Journal

    Article Title: Effects of calcitriol (1, 25-dihydroxy-vitamin D3) on the inflammatory response induced by H9N2 influenza virus infection in human lung A549 epithelial cells and in mice

    doi: 10.1186/s12985-017-0683-y

    Figure Lengend Snippet: The effects of calcitriol on A549 cells. a Effect of calcitriol treatment on virus growth kinetic in A549 cells. Cell culture supernatants were harvested every 24 h from infected cells and the viral load was titrated using the TCID50 assay. Real-time PCR was used to measure the effect of calcitriol treatment on the mRNA expression levels of the viral M gene ( b ), IL-6 ( c ), and IFN-β ( d ) in A549 cells. β-actin was used as an internal control. The data are expressed as mean ± SEM of triplicate samples

    Article Snippet: The effect of calcitriol on A549 cells A549 cells were grown in T-25 tissue culture flasks (Corning) to approximately 75% confluence prior to use in experiments.

    Techniques: Cell Culture, Infection, TCID50 Assay, Real-time Polymerase Chain Reaction, Expressing