a549 lung epithelial cells Search Results


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DSMZ a 549 human lung epithelial cells a 549
A 549 Human Lung Epithelial Cells A 549, supplied by DSMZ, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Corning Life Sciences a549 epithelial lung cells
The increased production of O2− by neutrophils in the presence of A. baumannii is associated with epithelial cell death. (A) Graphic representation of macrophages, neutrophils, and A. baumannii (Ab) interaction assay. Individual phagocytic cells or coculture and bacteria were coincubated in direct contact on a pore insert, placed on a microtiter well containing <t>A549</t> epithelial lung cells, and the supernatant collected after each interaction. (B) O2− dismutase activity was quantified in culture supernatants. The percentage of cytotoxicity among A549 cells was determined using the trypan blue exclusion test after coincubation with A. baumannii and phagocytic cells (C) or LPS (D). For panels B to D, each symbol (black, red, or blue circles and triangles) represents a technical replicate (B and C, n = 8; D, n = 6) per condition, dashed lines signify the average for each experimental condition, and error bars indicate SDs. Each experiment was performed independently twice, and the data obtained in each experiment were compiled into the graphs shown. For panel B, symbols (*, #, ϕ, P < 0.0001) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols *, #, and ϕ indicate higher O2− dismutase activity in supernatants of A549 cells incubated with A. baumannii plus neutrophils than in epithelial cells incubated with neutrophils, A. baumannii plus macrophages, and A. baumannii plus neutrophils plus macrophages, respectively. For panel C, symbols (*, #, ϕ, P < 0.0001; ϕ, P < 0.05) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols *, #, and ϕ indicate higher A549 cytotoxicity than A. baumannii, A. baumannii plus macrophages, and A. baumannii plus neutrophils plus macrophages, respectively. For panel D, symbols (*, #, P < 0.0001; *, P < 0.01) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols * and # indicate higher A549 cytotoxicity than in untreated and 1 μg/ml LPS-treated cells, respectively.
A549 Epithelial Lung Cells, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Thermo Fisher a549 epithelial lung cells
In vitro analysis of the interaction of C. neoformans alg3 Δ with host cells. (A) Analysis of adherence to lung epithelial cells. CFU of C. neoformans cells recovered from host cell lysates were counted after coincubation of C. neoformans cells with <t>A549</t> lung epithelial cells at 37°C with 5% CO 2 for 1 h. (B) Analysis of phagocytosis with or without opsonization. C. neoformans cells were incubated with activated J774A.1 macrophages for 2 h. CFU were counted to determine opsonic and nonopsonic phagocytosis. NS, not significant for all cells (determined by one-way analysis of variance and Bonferroni post hoc tests). (C) Analysis of phagosome acidification. Live and heat-killed C. neoformans cells were incubated with activated macrophage (J774A.1) for 2 h. Staining with LysoTracker Red was used to detect acidic phagosomes, and heat-killed C. neoformans cells served as a positive control for phagosomal acidification. The microscopy images, which overlap the confocal images, were obtained by d ifferential i nterference c ontrast (DIC). Representative images are presented with scale bars (20 μm). The percentage of acidified macrophages among Cryptococcus -containing phagolysosomes (CCPs) was calculated by counting the number of LysoTracker Red-stained phagolysosomes and dividing that number by the number of CCPs. Data representing means ± standard deviations were generated from three biologically independent experiments with at least 100 CCPs per infection. Statistical analysis was performed using an unpaired two-tailed Student's t test (ns, not significant; ** * , P < 0.0001).
A549 Epithelial Lung Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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BioResource International Inc lung epithelial cell line a 549
In vitro analysis of the interaction of C. neoformans alg3 Δ with host cells. (A) Analysis of adherence to lung epithelial cells. CFU of C. neoformans cells recovered from host cell lysates were counted after coincubation of C. neoformans cells with <t>A549</t> lung epithelial cells at 37°C with 5% CO 2 for 1 h. (B) Analysis of phagocytosis with or without opsonization. C. neoformans cells were incubated with activated J774A.1 macrophages for 2 h. CFU were counted to determine opsonic and nonopsonic phagocytosis. NS, not significant for all cells (determined by one-way analysis of variance and Bonferroni post hoc tests). (C) Analysis of phagosome acidification. Live and heat-killed C. neoformans cells were incubated with activated macrophage (J774A.1) for 2 h. Staining with LysoTracker Red was used to detect acidic phagosomes, and heat-killed C. neoformans cells served as a positive control for phagosomal acidification. The microscopy images, which overlap the confocal images, were obtained by d ifferential i nterference c ontrast (DIC). Representative images are presented with scale bars (20 μm). The percentage of acidified macrophages among Cryptococcus -containing phagolysosomes (CCPs) was calculated by counting the number of LysoTracker Red-stained phagolysosomes and dividing that number by the number of CCPs. Data representing means ± standard deviations were generated from three biologically independent experiments with at least 100 CCPs per infection. Statistical analysis was performed using an unpaired two-tailed Student's t test (ns, not significant; ** * , P < 0.0001).
Lung Epithelial Cell Line A 549, supplied by BioResource International Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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GE Healthcare lung epithelial a549 cells
Expression kinetics of endogenous interferon-inducible transmembrane protein 3 (IFITM3) protein modulated by virus infection. (A) <t>A549,</t> HeLa, and 293T cells were infected with VACV Tian Tan (VTT) at 0.1 PFU/cell for indicated time points after infection. Total cell proteins were extracted and the expression kinetics of IFITM3 and VTT D8 protein (*) were measured by Western blot and normalized to GAPDH. (B) Western blot analysis of IFITM1 and IFITM3 expression in HeLa cells treated with IFNα2b (10,000 U/mL) at indicated time point posttreatment. (C) Effect of IFNα2b at indicated concentrations on vaccinia virus infection in HeLa cells evaluated by flow cytometry and fluorescence microscope. (D,E) qPCR analysis of IFNα and IFNβ in HeLa and 293T cells infected by VTT infection at indicated time points postinfection. (F) Western blot analysis of IRF3 expression and phosphorylation in HeLa cells treated by VTT at indicated time points postinfection. ** P <0.01.
Lung Epithelial A549 Cells, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


The increased production of O2− by neutrophils in the presence of A. baumannii is associated with epithelial cell death. (A) Graphic representation of macrophages, neutrophils, and A. baumannii (Ab) interaction assay. Individual phagocytic cells or coculture and bacteria were coincubated in direct contact on a pore insert, placed on a microtiter well containing A549 epithelial lung cells, and the supernatant collected after each interaction. (B) O2− dismutase activity was quantified in culture supernatants. The percentage of cytotoxicity among A549 cells was determined using the trypan blue exclusion test after coincubation with A. baumannii and phagocytic cells (C) or LPS (D). For panels B to D, each symbol (black, red, or blue circles and triangles) represents a technical replicate (B and C, n = 8; D, n = 6) per condition, dashed lines signify the average for each experimental condition, and error bars indicate SDs. Each experiment was performed independently twice, and the data obtained in each experiment were compiled into the graphs shown. For panel B, symbols (*, #, ϕ, P < 0.0001) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols *, #, and ϕ indicate higher O2− dismutase activity in supernatants of A549 cells incubated with A. baumannii plus neutrophils than in epithelial cells incubated with neutrophils, A. baumannii plus macrophages, and A. baumannii plus neutrophils plus macrophages, respectively. For panel C, symbols (*, #, ϕ, P < 0.0001; ϕ, P < 0.05) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols *, #, and ϕ indicate higher A549 cytotoxicity than A. baumannii, A. baumannii plus macrophages, and A. baumannii plus neutrophils plus macrophages, respectively. For panel D, symbols (*, #, P < 0.0001; *, P < 0.01) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols * and # indicate higher A549 cytotoxicity than in untreated and 1 μg/ml LPS-treated cells, respectively.

Journal: Infection and Immunity

Article Title: Depletion of Alveolar Macrophages Increases Pulmonary Neutrophil Infiltration, Tissue Damage, and Sepsis in a Murine Model of Acinetobacter baumannii Pneumonia

doi: 10.1128/IAI.00128-20

Figure Lengend Snippet: The increased production of O2− by neutrophils in the presence of A. baumannii is associated with epithelial cell death. (A) Graphic representation of macrophages, neutrophils, and A. baumannii (Ab) interaction assay. Individual phagocytic cells or coculture and bacteria were coincubated in direct contact on a pore insert, placed on a microtiter well containing A549 epithelial lung cells, and the supernatant collected after each interaction. (B) O2− dismutase activity was quantified in culture supernatants. The percentage of cytotoxicity among A549 cells was determined using the trypan blue exclusion test after coincubation with A. baumannii and phagocytic cells (C) or LPS (D). For panels B to D, each symbol (black, red, or blue circles and triangles) represents a technical replicate (B and C, n = 8; D, n = 6) per condition, dashed lines signify the average for each experimental condition, and error bars indicate SDs. Each experiment was performed independently twice, and the data obtained in each experiment were compiled into the graphs shown. For panel B, symbols (*, #, ϕ, P < 0.0001) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols *, #, and ϕ indicate higher O2− dismutase activity in supernatants of A549 cells incubated with A. baumannii plus neutrophils than in epithelial cells incubated with neutrophils, A. baumannii plus macrophages, and A. baumannii plus neutrophils plus macrophages, respectively. For panel C, symbols (*, #, ϕ, P < 0.0001; ϕ, P < 0.05) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols *, #, and ϕ indicate higher A549 cytotoxicity than A. baumannii, A. baumannii plus macrophages, and A. baumannii plus neutrophils plus macrophages, respectively. For panel D, symbols (*, #, P < 0.0001; *, P < 0.01) denote P value significance calculated using ANOVA and adjusted by use of the Bonferroni correction. Symbols * and # indicate higher A549 cytotoxicity than in untreated and 1 μg/ml LPS-treated cells, respectively.

Article Snippet: Also, the activity of O 2 − dismutase was determined as described above after neutrophils (10 5 ), macrophages (10 5 ), or cocultures (10 5 cells per leukocyte) were incubated with A. baumannii (10 6 CFU) on an insert with a pore size of 0.4 μm (Corning) that was placed onto a 24-well polystyrene plate containing 10 5 A549 epithelial lung cells in feeding medium and incubated for 12 h at 37°C in 5% CO 2 ( ).

Techniques: Activity Assay, Incubation

In vitro analysis of the interaction of C. neoformans alg3 Δ with host cells. (A) Analysis of adherence to lung epithelial cells. CFU of C. neoformans cells recovered from host cell lysates were counted after coincubation of C. neoformans cells with A549 lung epithelial cells at 37°C with 5% CO 2 for 1 h. (B) Analysis of phagocytosis with or without opsonization. C. neoformans cells were incubated with activated J774A.1 macrophages for 2 h. CFU were counted to determine opsonic and nonopsonic phagocytosis. NS, not significant for all cells (determined by one-way analysis of variance and Bonferroni post hoc tests). (C) Analysis of phagosome acidification. Live and heat-killed C. neoformans cells were incubated with activated macrophage (J774A.1) for 2 h. Staining with LysoTracker Red was used to detect acidic phagosomes, and heat-killed C. neoformans cells served as a positive control for phagosomal acidification. The microscopy images, which overlap the confocal images, were obtained by d ifferential i nterference c ontrast (DIC). Representative images are presented with scale bars (20 μm). The percentage of acidified macrophages among Cryptococcus -containing phagolysosomes (CCPs) was calculated by counting the number of LysoTracker Red-stained phagolysosomes and dividing that number by the number of CCPs. Data representing means ± standard deviations were generated from three biologically independent experiments with at least 100 CCPs per infection. Statistical analysis was performed using an unpaired two-tailed Student's t test (ns, not significant; ** * , P < 0.0001).

Journal: mBio

Article Title: Core N -Glycan Structures Are Critical for the Pathogenicity of Cryptococcus neoformans by Modulating Host Cell Death

doi: 10.1128/mBio.00711-20

Figure Lengend Snippet: In vitro analysis of the interaction of C. neoformans alg3 Δ with host cells. (A) Analysis of adherence to lung epithelial cells. CFU of C. neoformans cells recovered from host cell lysates were counted after coincubation of C. neoformans cells with A549 lung epithelial cells at 37°C with 5% CO 2 for 1 h. (B) Analysis of phagocytosis with or without opsonization. C. neoformans cells were incubated with activated J774A.1 macrophages for 2 h. CFU were counted to determine opsonic and nonopsonic phagocytosis. NS, not significant for all cells (determined by one-way analysis of variance and Bonferroni post hoc tests). (C) Analysis of phagosome acidification. Live and heat-killed C. neoformans cells were incubated with activated macrophage (J774A.1) for 2 h. Staining with LysoTracker Red was used to detect acidic phagosomes, and heat-killed C. neoformans cells served as a positive control for phagosomal acidification. The microscopy images, which overlap the confocal images, were obtained by d ifferential i nterference c ontrast (DIC). Representative images are presented with scale bars (20 μm). The percentage of acidified macrophages among Cryptococcus -containing phagolysosomes (CCPs) was calculated by counting the number of LysoTracker Red-stained phagolysosomes and dividing that number by the number of CCPs. Data representing means ± standard deviations were generated from three biologically independent experiments with at least 100 CCPs per infection. Statistical analysis was performed using an unpaired two-tailed Student's t test (ns, not significant; ** * , P < 0.0001).

Article Snippet: To analyze the adhesion of yeast cells to (A549) epithelial lung cells, 2 × 10 5 A549 cells were seeded into 24-well plates in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) supplemented with 10% fetal bovine serum (FBS) and were cultivated at 37°C in 5% CO 2 for 18 h. C. neoformans cells (2 × 10 6 ) were added to each well ( C. neoformans /A549 ratio, 10:1) and coincubated at 37°C for 1 h. Then, each well was gently washed three times with PBS to remove nonadherent yeast cells.

Techniques: In Vitro, Incubation, Staining, Positive Control, Microscopy, Generated, Infection, Two Tailed Test

Expression kinetics of endogenous interferon-inducible transmembrane protein 3 (IFITM3) protein modulated by virus infection. (A) A549, HeLa, and 293T cells were infected with VACV Tian Tan (VTT) at 0.1 PFU/cell for indicated time points after infection. Total cell proteins were extracted and the expression kinetics of IFITM3 and VTT D8 protein (*) were measured by Western blot and normalized to GAPDH. (B) Western blot analysis of IFITM1 and IFITM3 expression in HeLa cells treated with IFNα2b (10,000 U/mL) at indicated time point posttreatment. (C) Effect of IFNα2b at indicated concentrations on vaccinia virus infection in HeLa cells evaluated by flow cytometry and fluorescence microscope. (D,E) qPCR analysis of IFNα and IFNβ in HeLa and 293T cells infected by VTT infection at indicated time points postinfection. (F) Western blot analysis of IRF3 expression and phosphorylation in HeLa cells treated by VTT at indicated time points postinfection. ** P <0.01.

Journal: Frontiers in Immunology

Article Title: The Host Restriction Factor Interferon-Inducible Transmembrane Protein 3 Inhibits Vaccinia Virus Infection

doi: 10.3389/fimmu.2018.00228

Figure Lengend Snippet: Expression kinetics of endogenous interferon-inducible transmembrane protein 3 (IFITM3) protein modulated by virus infection. (A) A549, HeLa, and 293T cells were infected with VACV Tian Tan (VTT) at 0.1 PFU/cell for indicated time points after infection. Total cell proteins were extracted and the expression kinetics of IFITM3 and VTT D8 protein (*) were measured by Western blot and normalized to GAPDH. (B) Western blot analysis of IFITM1 and IFITM3 expression in HeLa cells treated with IFNα2b (10,000 U/mL) at indicated time point posttreatment. (C) Effect of IFNα2b at indicated concentrations on vaccinia virus infection in HeLa cells evaluated by flow cytometry and fluorescence microscope. (D,E) qPCR analysis of IFNα and IFNβ in HeLa and 293T cells infected by VTT infection at indicated time points postinfection. (F) Western blot analysis of IRF3 expression and phosphorylation in HeLa cells treated by VTT at indicated time points postinfection. ** P <0.01.

Article Snippet: Human embryonic kidney cells (HEK-293T), baby hamster kidney cells (BHK-21), African green monkey kidney epithelial cells (Vero), human cervical carcinoma cells (HeLa), and lung epithelial (A549) cells were grown in complete DMEM (HyClone) with 10% fetal bovine serum (Thermo) at 37°C in a 5% CO 2 incubator.

Techniques: Expressing, Infection, Western Blot, Flow Cytometry, Fluorescence, Microscopy

Interferon-inducible transmembrane protein 3 (IFITM3) silencing mediates antiviral actions of IFN-α2b. (A,C) IFITM3 expression was analyzed by Western blot in siRNA-transfected HeLa and 293T cells treated with or without IFN-α2b. (B,D) Green fluorescent protein (EGFP)-positive cells were scored by flow cytometry of siRNA-transfected HeLa and 293T cells with or without IFN-α2b treatment at 24 h postinfection by vaccinia virus Tian Tan green fluorescent protein (VTT-EGFP). (E,F) Viability of siRNA-transfected HeLa and A549 cells infected with VTT virus at 0.1 PFU/cell was assessed by MTS assay. Values represent the mean ± SD, n = 4. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Journal: Frontiers in Immunology

Article Title: The Host Restriction Factor Interferon-Inducible Transmembrane Protein 3 Inhibits Vaccinia Virus Infection

doi: 10.3389/fimmu.2018.00228

Figure Lengend Snippet: Interferon-inducible transmembrane protein 3 (IFITM3) silencing mediates antiviral actions of IFN-α2b. (A,C) IFITM3 expression was analyzed by Western blot in siRNA-transfected HeLa and 293T cells treated with or without IFN-α2b. (B,D) Green fluorescent protein (EGFP)-positive cells were scored by flow cytometry of siRNA-transfected HeLa and 293T cells with or without IFN-α2b treatment at 24 h postinfection by vaccinia virus Tian Tan green fluorescent protein (VTT-EGFP). (E,F) Viability of siRNA-transfected HeLa and A549 cells infected with VTT virus at 0.1 PFU/cell was assessed by MTS assay. Values represent the mean ± SD, n = 4. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Human embryonic kidney cells (HEK-293T), baby hamster kidney cells (BHK-21), African green monkey kidney epithelial cells (Vero), human cervical carcinoma cells (HeLa), and lung epithelial (A549) cells were grown in complete DMEM (HyClone) with 10% fetal bovine serum (Thermo) at 37°C in a 5% CO 2 incubator.

Techniques: Expressing, Western Blot, Transfection, Flow Cytometry, Infection, MTS Assay

Interferon-inducible transmembrane protein 3 (IFITM3) overexpression affects pH-dependent vaccinia virus (VACV) entry. (A,B) IFITM3 + 293T and baby hamster kidney cells were infected with vaccinia virus Tian Tan green fluorescent protein (VTT-EGFP) at the indicated MOI on ice for 1 h, followed by either mock treatment or treatment with 10 or 20 mM NH 4 Cl at 37°C for 24 h. Percentages of infected cells were determined by flow cytometry. (C) Plaque forming assay of Vero cell lines, which were incubated with VTT at 0.01 MOI on ice for 1 h and then treated with 10 mM NH 4 Cl or buffer (mock) at 37°C for 48 h. (D–F) A549 or 293T cells were incubated with VTT-EGFP at the indicated MOI on ice for 1 h and then treated with 10 mM NH 4 Cl or PBS buffer (mock) (D) and pH 5.0 buffer, pH 4.0 buffer, or PBS (mock) (E,F) at 37°C for 24 h. Percentages of infected cells were determined by flow cytometry. All values represent the mean ± SD, n = 3. * P < 0.05 and ** P < 0.01.

Journal: Frontiers in Immunology

Article Title: The Host Restriction Factor Interferon-Inducible Transmembrane Protein 3 Inhibits Vaccinia Virus Infection

doi: 10.3389/fimmu.2018.00228

Figure Lengend Snippet: Interferon-inducible transmembrane protein 3 (IFITM3) overexpression affects pH-dependent vaccinia virus (VACV) entry. (A,B) IFITM3 + 293T and baby hamster kidney cells were infected with vaccinia virus Tian Tan green fluorescent protein (VTT-EGFP) at the indicated MOI on ice for 1 h, followed by either mock treatment or treatment with 10 or 20 mM NH 4 Cl at 37°C for 24 h. Percentages of infected cells were determined by flow cytometry. (C) Plaque forming assay of Vero cell lines, which were incubated with VTT at 0.01 MOI on ice for 1 h and then treated with 10 mM NH 4 Cl or buffer (mock) at 37°C for 48 h. (D–F) A549 or 293T cells were incubated with VTT-EGFP at the indicated MOI on ice for 1 h and then treated with 10 mM NH 4 Cl or PBS buffer (mock) (D) and pH 5.0 buffer, pH 4.0 buffer, or PBS (mock) (E,F) at 37°C for 24 h. Percentages of infected cells were determined by flow cytometry. All values represent the mean ± SD, n = 3. * P < 0.05 and ** P < 0.01.

Article Snippet: Human embryonic kidney cells (HEK-293T), baby hamster kidney cells (BHK-21), African green monkey kidney epithelial cells (Vero), human cervical carcinoma cells (HeLa), and lung epithelial (A549) cells were grown in complete DMEM (HyClone) with 10% fetal bovine serum (Thermo) at 37°C in a 5% CO 2 incubator.

Techniques: Over Expression, Infection, Flow Cytometry, Incubation