human epithelial lung cell line a549  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Nebulized Hybrid Nanoarchaeosomes: Anti-Inflammatory Activity, Anti-Microbial Activity and Cytotoxicity on A549 Cells

doi: 10.3390/ijms26010392

Figure Lengend Snippet: IC 50 values of formulations on A549 cells.

Article Snippet: Human epithelial lung cell line A549 (ATCC ® CCL-185™) was maintained in MEM supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Nebulized Hybrid Nanoarchaeosomes: Anti-Inflammatory Activity, Anti-Microbial Activity and Cytotoxicity on A549 Cells

doi: 10.3390/ijms26010392

Figure Lengend Snippet: A549 cell viability after 72 h when treated with ( A ) BSs and ( B ) hybrid-nanoARCs. Statistical significance compared with medium control was determined using a Kruskal– Wallis non-parametric test followed by Dunn’s multiple comparisons, * p < 0.05, ** p < 0.01; **** p < 0.0001.

Article Snippet: Human epithelial lung cell line A549 (ATCC ® CCL-185™) was maintained in MEM supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Nebulized Hybrid Nanoarchaeosomes: Anti-Inflammatory Activity, Anti-Microbial Activity and Cytotoxicity on A549 Cells

doi: 10.3390/ijms26010392

Figure Lengend Snippet: A549 cell viability after 72 h when treated with nebulized BSs and BS + BS-nanoARCs. Statistical significance compared with medium control was determined using a Kruskal– Wallis non-parametric test followed by Dunn’s multiple comparisons, ** p < 0.01; **** p < 0.0001.

Article Snippet: Human epithelial lung cell line A549 (ATCC ® CCL-185™) was maintained in MEM supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Nebulized Hybrid Nanoarchaeosomes: Anti-Inflammatory Activity, Anti-Microbial Activity and Cytotoxicity on A549 Cells

doi: 10.3390/ijms26010392

Figure Lengend Snippet: Stability upon storage: ( A1 ) plasmonic peak and ( A2 ) size of BSs. A549 cell viability after treatment with stored ( B ) BSs and ( C ) [BS + BS-nanoARCs] (50 μg PL/mL, 12.5–25 μg total Ag/mL). Statistical significance compared with medium control was determined using a Kruskal–Wallis non-parametric test followed by Dunn’s multiple comparisons, ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human epithelial lung cell line A549 (ATCC ® CCL-185™) was maintained in MEM supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine.

Techniques: Control

Journal: mBio

Article Title: Naturally occurring influenza reassortment in pigs facilitates the emergence of intrahost virus subpopulations with distinct genotypes and replicative fitness

doi: 10.1128/mbio.01924-24

Figure Lengend Snippet: Growth performance of influenza A viral plaques with distinct genotypes on various human and swine respiratory epithelial cell monolayers. ( A ) Influenza growth kinetics of plaque-purified isolates on the normal swine bronchial epithelial cell (NSBE), swine nasal primary epithelial cell (sNEC), human bronchial/tracheal epithelial cell (NHBE), human nasal primary epithelial cell (HNEpC), and human lung carcinoma epithelial cell (A549) grown as monolayers. The cells were infected by indicated viruses at 33°C, 37°C, or 39°C with a multiplicity of infection of 0.01 TCID 50 /cell. The supernatants of infected cells were collected at 1, 24, 48, and 72 hours postinoculation (hpi), and the virus titers were determined by TCID 50 assay. The tested influenza viruses are shown in different colors, and the data are shown as mean titers ± standard errors of three replicates. ( B ) The 21 influenza viral plaques representing 21 genotypes were inoculated on NSBE, sNEC, NHBE, HNEpC, and A549 cells at a multiplicity of infection of 0.01 TCID 50 /cell and were cultured at either 33°C, 37°C, or 39°C. We computed the area under the curve (AUC) for each virus based on the virus growth kinetic curves shown in . The AUC values of each viral plaque are displayed as the mean values ± standard deviations ( n = 3 replicates). Each cell line is represented by a different color. ( C ) Statistical differences compared the average AUC values among all the 21 influenza genotypes for each cell line using two-way ANOVA with Tukey HSD for multiple group comparisons. The X and Y axes indicate the ID of the plaques with (Y label) or without (X label) the genotypes they represented from that used for each pairwise comparison. The colored squares indicate statistical differences ( P < 0.05) of the average AUC values between two viruses. The colored squares with pink borders indicate the significant differences in AUC values of two viral plaques/genotypes isolated from the same pig.

Article Snippet: The human lung carcinoma epithelial cell line (A549, CCL-185) was purchased from ATCC and cultured in DMEM media with 10% FBS and 1% streptomycin-penicillin.

Techniques: Purification, Infection, Virus, Cell Culture, Comparison, Isolation

Journal: mBio

Article Title: Naturally occurring influenza reassortment in pigs facilitates the emergence of intrahost virus subpopulations with distinct genotypes and replicative fitness

doi: 10.1128/mbio.01924-24

Figure Lengend Snippet: Growth performance of influenza plaques with distinct genotypes tested on swine and human respiratory primary epithelial cells at the air–liquid interface. ( A ) Influenza growth kinetics of selected plaque-purified isolates on human bronchial/tracheal epithelial (NHBE-ALI) and swine tracheal primary epithelial (sTEC-ALI) cells at the air–liquid interface. The selected representative influenza viruses were inoculated on the cells at 37°C with a multiplicity of infection of 0.1 TCID 50 /cell. The apical supernatants were obtained at 3, 24, 48, and 72 hours postinoculation (hpi), and the viral titers were determined with TCID 50 assay. The influenza viruses are displayed in different colors and shapes. Error bars present the standard error of the means from three replicates. ( B ) The NHBE-ALI and the sTEC-ALI cells were cultured and inoculated in 37°C under air–liquid interface conditions with 23 influenza plaque viruses representing 21 genotypes at a multiplicity of infection of 0.1 TCID 50 /cell. The area under the curve (AUC) was calculated based on the virus growth kinetic curves shown in . ( C ) Statistical analysis on the average AUC values among the 23 influenza plaques tested on each cell line using two-way ANOVA with Tukey HSD adjustments. The figure settings of panels A, B, and C are the same as the corresponding figure panels in .

Article Snippet: The human lung carcinoma epithelial cell line (A549, CCL-185) was purchased from ATCC and cultured in DMEM media with 10% FBS and 1% streptomycin-penicillin.

Techniques: Purification, Infection, Cell Culture, Virus

Journal: PNAS Nexus

Article Title: Physiologic and structural characterization of desisobutyryl-ciclesonide, a selective glucocorticoid receptor modulator in newborn rats

doi: 10.1093/pnasnexus/pgae573

Figure Lengend Snippet: DEX and Des-CIC suppression of proinflammatory cytokine gene expression. A) Paradigm for neonatal rats treated daily with VEH, DEX (0.1 mg/kg), or Des-CIC (1.25 mg/kg) s.c. before i.p. injection of bleomycin (BLEO) (1 mg/kg) for 11 consecutive days. Relative mRNA expression levels for B) Il-1β , C) Il-6 , and D) Tnfα, compared to Actb in lung tissue of treated animals ( n = 7–9 per group). All data are expressed as mean ± SEM. Relative mRNA expression levels for IL-1β (E), IL-6 (F), and IL-8 (G) compared to glyceraldehyde-3-phosphate dehydrogenase in A549 cells exposed to various treatments ( n = 5 per group). All data are expressed as mean ± SEM. For data in (B–G), statistical significance was determined by Brown–Forsythe and Welch ANOVA tests with Dunnett's T3 post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. (H–J) Naïve CD8 + T cells from C57BL/6 mice were differentiated into effector-like cells in the presence of vehicle (DMSO), 1 µM DEX, or 1 µM Des-CIC for the final 24 h in culture. T cells were analyzed for the expression of IFNγ and TNFα from single, live cells by flow cytometry. Flow data from a representative experiment are shown in (H) with summary data shown in (I) and (J) for IFNγ and TNFα, respectively ( n = 5 per group). Data in (I) and (J) are expressed in mean ± SEM. Statistical significance was determined by ANOVA followed by Dunnett's multiple comparison test, **** P < 0.0001.

Article Snippet: Human lung epithelial A549 cells (ATCC) were plated at 250,000 in six well plates (Corning Costar) in Dulbecco’s Modified Eagle Medium/F12 containing L- glutamine (Corning) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals) and 50 units/mL Penicillin (Sigma) and 0.5 mg/mL streptomycin (Sigma).

Techniques: Expressing, Injection, Flow Cytometry, Comparison

Journal: International Journal of Biological Sciences

Article Title: The Cyclin-Dependent Kinase 8 Inhibitor E966-0530-45418 Attenuates Pulmonary Fibrosis In Vitro and In Vivo

doi: 10.7150/ijbs.105826

Figure Lengend Snippet: E966-0530-45418 is not cytotoxic and does not affect cell cycle progression. ( A ) The structure of E966-0530-45418. ( B ) Molecular docking analysis showed that E966-0530-45418 (gray) shows favorable interactions within the binding site in CDK8 (blue). The docking pose is represented as sticks. Binding site residues are rendered as lines and labeled. Halogen interactions and hydrogen bonds are denoted as purple or green lines, respectively. ( C - F ) Cell viability was measured by MTT assay for A549 (C), WI-38 (D), and THP-1 (E) cells and human primary alveolar epithelial cells (AECs) (F), which were incubated with different concentrations (0, 1, 2.5, 5, 10, or 20 μM) of E966-0530-45418 for 12, 24 or 48 h (n = 5 independent samples per group). The IC 50 values were calculated by a sigmoidal dose-response equation. The results are presented as the mean, along with the individual replicates. ( G ) Flow cytometric analysis of PI staining was used to evaluate the cell cycle distribution of human primary AECs treated with or without different concentrations (1, 3, or 10 μM) of E966-0530-45418 for 12, 24, or 48 h (n = 3 independent samples per group). The results are shown as the mean ± SEM. No significant statistics were determined using two-way ANOVA.

Article Snippet: The human lung epithelial cell line A549, human lung fibroblast cell line WI-38, and monocyte cell line THP-1 were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Binding Assay, Labeling, MTT Assay, Incubation, Staining

Journal: International Journal of Biological Sciences

Article Title: The Cyclin-Dependent Kinase 8 Inhibitor E966-0530-45418 Attenuates Pulmonary Fibrosis In Vitro and In Vivo

doi: 10.7150/ijbs.105826

Figure Lengend Snippet: E966-0530-45418 significantly attenuates the TGFβ1-induced increase of EMT proteins, fibrotic markers, and cell migration. ( A - J ) A549 cells were exposed to the indicated concentrations of E966-0530-45418 (μM), senexin A (5 μM), pirfenidone (1 mM), or no inhibitor in the presence of TGFβ1 (10 ng/mL) for 24 h. The protein levels of E-cadherin, N-cadherin, snail, α-SMA, and COL1A1 were determined by western blot in A549 cells (n = 4 independent samples per group, except for COL1A1, where n = 6) (A-D). The mRNA levels of E-cadherin, snail, α-SMA, COL1A1, TGFβ1, and CTGF were analyzed by RT‒qPCR in A549 cells (n = 3 independent samples per group, except for COL1A1, where n = 5) (E‒J). ( K , L ) A549 cells were treated with E966-0530-45418 at the indicated concentrations (μM), senexin A (5 μM), pirfenidone (1 mM), or no inhibitor in the presence of TGFβ1 (10 ng/mL), allowed to migrate into the wound area for 24 h and photographed; the cyan solid line depicts the edge between the cell-occupying region and the wound area (40× magnification) (Scale bar: 100 μm) (K). Cell migration into the wound was quantified using ImageJ software (L). (n = 4 independent samples per group). The results are shown as the mean ± SEM. P values were determined using one-way ANOVA followed by Tukey's post hoc test (B-J, and L).

Article Snippet: The human lung epithelial cell line A549, human lung fibroblast cell line WI-38, and monocyte cell line THP-1 were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Migration, Western Blot, Software

Journal: International Journal of Biological Sciences

Article Title: The Cyclin-Dependent Kinase 8 Inhibitor E966-0530-45418 Attenuates Pulmonary Fibrosis In Vitro and In Vivo

doi: 10.7150/ijbs.105826

Figure Lengend Snippet: E966-0530-45418 markedly mitigated TGFβ1/Smad3/RNA polymerase II signal transduction, subsequently lowering the transcription of EMT and fibrosis-related genes. ( A - C , E - G ) A549 cells were treated with E966-0530-45418 (5 μM), senexin A (5 μM), pirfenidone (1 mM), or no inhibitor in the presence of TGFβ1 (10 ng/mL) for 3 h and then subjected to nuclear-cytosolic fractionation. The protein levels in the cytosol and nucleus were detected by western blotting and quantified (n = 5 independent samples per group) (A-C). The nuclear proteins were immunoprecipitated with an anti-CDK8 antibody and then subjected to immunoblotting to assess the interactions between CDK8 and p-Smad3 T179, p-RNA Pol II S2/5, MED12, Pin 1, and cyclin C (n = 4 independent samples per group) (E-G). ( D ) Protein-protein interaction (PPI) network analysis was established by the STRING online database. The network nodes represent proteins, and the colored edges denote evidence of interactions between different proteins in the PPI network. Strong connections and networks were clustered using the k-means cluster algorithm with default parameters and are indicated by solid lines. POLR2A: RNA polymerase II. ( H ) Schematic illustrating Smad3 binding to the Smad-binding element (sequence logo obtained from the UCSC JASPAR joint website). ( I - K ) A549 cells were incubated with E966-0530-45418 (5 μM), senexin A (5 μM), pirfenidone (1 mM), or no inhibitor in the presence of TGFβ1 (10 ng/mL) for 6 h and then subjected to a Smad3 ChIP assay with RT‒qPCR analysis of the promoter sequences of N-cadherin, Snail (I), COL1A1, α-SMA (J), TGFβ1 and CTGF (K). The histone H3 antibody (Ab) group was used as a positive control, and the rabbit IgG group was used as a negative control (n = 3 independent samples per group). ( L ) Schematic illustration of the human E-cadherin and TGFβ1 promoter luciferase reporter genes, the function of which might be affected by TGFβ1 treatment. ( M ) A549 cells were transfected with 7TFP CDH1 or pGL3- TGFB1 reporter plasmid (1 μg) for 6 h, followed by subsequent treatment with TGF-β1 (10 ng/mL) with E966-0530-45418 (5 µM), senexin A (5 µM), pirfenidone (1 mM), or no inhibitor for 24 h, and luciferase expression was subsequently determined (n = 5 independent samples per group). The results are shown as the mean ± SEM. P values were determined using one-way ANOVA followed by Tukey's post hoc test (B, C, F, G, I-K, and M).

Article Snippet: The human lung epithelial cell line A549, human lung fibroblast cell line WI-38, and monocyte cell line THP-1 were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Transduction, Fractionation, Western Blot, Immunoprecipitation, Binding Assay, Sequencing, Incubation, Positive Control, Negative Control, Luciferase, Transfection, Plasmid Preparation, Expressing

Journal: Nature Communications

Article Title: A coopetition-driven strategy of parallel/perpendicular aromatic stacking enabling metastable supramolecular polymerization

doi: 10.1038/s41467-024-55106-z

Figure Lengend Snippet: a Fluorescence spectral variations of 1AggI ( c = 0.80 × 10 –5 M) in cell culture medium with 0.05% DMSO. b The corresponding kinetic profiles of the conversion from 1AggI to 1AggII by monitoring the emission intensities at 550 and 600 nm. λ ex = 405 nm. CLSM images of the spontaneous transformation of 1AggI to 1AggII in A549 cells at ( c – e ) red channel ( λ ex = 405 nm, λ em = 600–700 nm) and ( f – h ) green channel ( λ ex = 405 nm, λ em = 500–599 nm) for 0, 15, and 30 min. Scale bar: 40 μm. Cell imaging was repeated at least three times with similar results.

Article Snippet: The human lung adenocarcinoma epithelial cell line A549 (male, CCL-185) was obtained from the Chinese Academy of Science Cell Bank, and B16-F10 cell line (ATCC, CRL-6475) was purchased from Wuhan Pricella Biotechnology Co., Ltd.

Techniques: Fluorescence, Cell Culture, Transformation Assay, Imaging

Journal: BMC Microbiology

Article Title: An improvised one-step OptiPrep cushion ultracentrifugation method for outer membrane vesicles isolation of Klebsiella pneumoniae

doi: 10.1186/s12866-024-03649-y

Figure Lengend Snippet: Differences in biological activity between K. pneumoniae -OMVs extracted by DC and DDGC. ( A ) LDH release assay to evaluate the cytotoxicity of K. pneumoniae -OMVs: OMVs extracted by the DDGC method showed higher cytotoxicity at concentrations of 20 µg/mL and 50 µg/mL compared to those extracted by the DC method (** P < 0.01); ( B ) Analysis of A549 cell toxicity induced by K. pneumoniae -OMVs: The impact on A549 cell vitality was significant at 20ug/mL and 50ug/mL, with a notable difference between the two extraction methods (** P < 0.01). At a concentration of 100 µg/mL of K. pneumoniae -OMVs, significant cytotoxicity was observed, with cell survival rates induced by OMVs extracted by both methods falling below 40%; ( C - F ) Impact of K. pneumoniae -OMVs on the RT-qPCR Expression of Inflammatory Factors in A549 Cells. The activation effect of OMVs extracted by DC and DDGC at concentrations of 20 µg/mL and 50 µg/mL on (C) IL-6 , ( D ) TNF-α , ( E ) IL-1β , and ( F ) IL-8 mRNA levels. LPS served as a positive control, and untreated cells as a negative control (NC). The activation effect increased with concentration, with OMVs extracted by DDGC being more effective than those extracted by DC, especially at 50 µg/mL. Statistical significance is indicated by asterisks (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The A549 Human Lung Epithelial Cell Line (FH0045) were obtained from FuHeng Cell Center (Shanghai, China) and maintained in DMEM medium (Gibco, United States) supplemented with 10% fetal bovine serum (FBS, Gibco, United States) under a humidified atmosphere of 5% CO2 at 37 ◦C.

Techniques: Activity Assay, Lactate Dehydrogenase Assay, Extraction, Concentration Assay, Quantitative RT-PCR, Expressing, Activation Assay, Positive Control, Negative Control