human lung carcinoma epithelial cell line  (ATCC)

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: Pharmaceuticals

Article Title: Stephania tetrandra and Its Active Compound Coclaurine Sensitize NSCLC Cells to Cisplatin through EFHD2 Inhibition

doi: 10.3390/ph17101356

Figure Lengend Snippet: Characterization of coclaurine in EFHD2 inhibition and cancer biology. ( A ) Cell viability of H1299 and A549 cells treated the indicated doses of coclaurine was analyzed by MTT assay. ( B ) The expression of EFHD2 and its related signaling molecules in H1299 and A549 cells treated or untreated with coclaurine (200 μM) were determined by Western blot. ( C ) The intracellular ROS levels in H1299 and A549 cells treated or untreated with coclaurine were determined using the tracer dye CM-H2DCFDA (DCF, Invitrogen). ( D ) Cell viability of H1299 and A549 cells treated or untreated with coclaurine (200 μM) at the indicated doses of cisplatin was analyzed by MTT assay. ( E ) Colony formation assay in H1299 and A549 cells and ( F , G ) cell migration assay and invasion assay in H1299 cells treated with the indicated doses of coclaurine. Signal quantification using crystal violet extract was measured by colorimetric analysis at 570 nm. The relative signal intensities were normalized to the untreated control ( n = 3). Scale bar, 0.1 cm. ( H ) H1299 and A549 cells were treated with the indicated doses of coclaurine for 10 d, the number of spheroid formation was counted manually under a bright-field microscope ( n = 3). Scale bar, 100 μm. ( I ) The expression of stemness-related proteins in H1299 and A549 cells treated or untreated with coclaurine (200 μM) was determined by Western blot. β-actin, loading control. Data are displayed as the means ± SD. For statistical analysis, a 2-tailed unpaired Student’s t -test ( C , E – H ). ★ , p < 0.05; ★★ , p < 0.01.

Article Snippet: Human lung epithelial carcinoma cell lines A549 and H1299, as well as the human non-tumorigenic bronchial epithelial cell line BEAS-2B, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Inhibition, MTT Assay, Expressing, Western Blot, Colony Assay, Cell Migration Assay, Invasion Assay, Control, Microscopy

Journal: Pharmaceuticals

Article Title: Stephania tetrandra and Its Active Compound Coclaurine Sensitize NSCLC Cells to Cisplatin through EFHD2 Inhibition

doi: 10.3390/ph17101356

Figure Lengend Snippet: Coclaurine suppresses EFHD2 through inhibition of the transcriptional activity of FOXG1. ( A ) After cycloheximide (CHX, 10 mg/mL) treatment for 1 h, H1299 and A549 cells were treated or untreated with coclaurine (200 μM) for the indicated times. EFHD2 expression in each experimental group was determined by Western blot. ( B ) EFHD2 mRNA levels of H1299 and A549 cells treated or untreated with coclaurine (200 μM) for 24 h were determined by qPCR. GADHP and 18S rRNA served as internal controls for gene expression. ( C ) Schematic illustration showing the construction of the EFHD2 promoter sequence in the luciferase expression vector. ( D ) Relative luciferase activity with various EFHD2 promoter sequences in H1299 cells treated with coclaurine (200 μM) compared to the untreated control. ( E ) Schematic illustration showing the binding sites of transcription factors and the alteration of CDOCKER energy by coclaurine binding. ( F , G ) Three- and two-dimensional interactions between transcription factor FOXG1 (green), DNA (gray), and coclaurine (red) in two potential binding pockets. The binding amino acids in FOXG1 and DNA nucleotides with coclaurine are shown in the two-dimensional structures. ( H ) FOXG1 thermal stability in H1299 cells treated or untreated with coclaurine (200 μM) at the indicated temperatures was determined by Western blot. The relative FOXG1 level (FOXG1/actin) was normalized to the unheated control of each group. ( I ) FOXG1 and EFHD2 expression and ( J ) cell viability of H1299 cells with or without FOXG1/KD in the presence or absence of coclaurine (200 μM) were determined by Western blot and MTT assay, respectively. Data are displayed as the means ± SD. For statistical analysis, a 2-tailed unpaired Student’s t -test ( J ). ★★ , p < 0.01.

Article Snippet: Human lung epithelial carcinoma cell lines A549 and H1299, as well as the human non-tumorigenic bronchial epithelial cell line BEAS-2B, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Inhibition, Activity Assay, Expressing, Western Blot, Sequencing, Luciferase, Plasmid Preparation, Control, Binding Assay, MTT Assay

Journal: International Journal of Molecular Sciences

Article Title: Staphylococcus aureus Co-Infection in COVID-19 Patients: Virulence Genes and Their Influence on Respiratory Epithelial Cells in Light of Risk of Severe Secondary Infection

doi: 10.3390/ijms251810050

Figure Lengend Snippet: Adhesion of S. aureus strains measured by % of recovered bacteria not adhered to respiratory epithelial cells (A549 line). n/n′—ratio of counts per 60 s of sample flow before and after exposition to epithelial cells (A549). The circle represents the result (median) of a single strain; the size of the circle represents the relative initial inoculum of the sample.

Article Snippet: The A549 human lung carcinoma epithelial cell line (ATCC, CCL-185) (ATCC, Manassas, VA, USA) was cultured in T75 bottles (Googlab Scientific, Rokocin, Poland) in a F-12K culture medium (ATCC, Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin (all from Merck, Darmstadt, Germany) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 in the Thermo Scientific Forma Steri-Cycle CO 2 Incubator (Thermo Fisher Scientific, Waltham, MA, USA) [ , ].

Techniques: Bacteria

Journal: International Journal of Molecular Sciences

Article Title: Staphylococcus aureus Co-Infection in COVID-19 Patients: Virulence Genes and Their Influence on Respiratory Epithelial Cells in Light of Risk of Severe Secondary Infection

doi: 10.3390/ijms251810050

Figure Lengend Snippet: Adhesion of the S. aureus cell line in relation to the presence of adhesin genes. % of recovery of unattached A549 bacterial cells to the initial bacterial suspension. The vertical bars represent standard errors. The statistical analysis was performed using the Kruskal–Wallis test.

Article Snippet: The A549 human lung carcinoma epithelial cell line (ATCC, CCL-185) (ATCC, Manassas, VA, USA) was cultured in T75 bottles (Googlab Scientific, Rokocin, Poland) in a F-12K culture medium (ATCC, Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin (all from Merck, Darmstadt, Germany) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 in the Thermo Scientific Forma Steri-Cycle CO 2 Incubator (Thermo Fisher Scientific, Waltham, MA, USA) [ , ].

Techniques: Suspension

Journal: International Journal of Molecular Sciences

Article Title: Staphylococcus aureus Co-Infection in COVID-19 Patients: Virulence Genes and Their Influence on Respiratory Epithelial Cells in Light of Risk of Severe Secondary Infection

doi: 10.3390/ijms251810050

Figure Lengend Snippet: The percentage of COVID-19 (black numbers) and non-COVID-19 (red numbers) S. aureus cells recovered from the inside of A549 pulmonary epithelial cells. The arithmetic mean of the triplicates, including the error bars that represents the SD. The statistical analysis was performed using a t -test. No statistical differences were found between the strains recovered from the inside of A549 cells.

Article Snippet: The A549 human lung carcinoma epithelial cell line (ATCC, CCL-185) (ATCC, Manassas, VA, USA) was cultured in T75 bottles (Googlab Scientific, Rokocin, Poland) in a F-12K culture medium (ATCC, Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin (all from Merck, Darmstadt, Germany) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 in the Thermo Scientific Forma Steri-Cycle CO 2 Incubator (Thermo Fisher Scientific, Waltham, MA, USA) [ , ].

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Staphylococcus aureus Co-Infection in COVID-19 Patients: Virulence Genes and Their Influence on Respiratory Epithelial Cells in Light of Risk of Severe Secondary Infection

doi: 10.3390/ijms251810050

Figure Lengend Snippet: The cell viability and LDH release assay for the A549 cells infected with different S. aureus strains (COVID-19 strains are marked in black, and non-COVID-19 strains are marked in red) after 4 h of incubation compared to the uninfected cells and the lysis-buffer-treated positive control. The mean values from the three independent experiments are shown, with the error bars representing the SD. The statistical analysis was performed using the Kruskal–Wallis test, followed by Dunn’s multiple comparison test for the values with a nonparametric distribution, with *** p < 0.001 and ** p < 0.01.

Article Snippet: The A549 human lung carcinoma epithelial cell line (ATCC, CCL-185) (ATCC, Manassas, VA, USA) was cultured in T75 bottles (Googlab Scientific, Rokocin, Poland) in a F-12K culture medium (ATCC, Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin (all from Merck, Darmstadt, Germany) at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 in the Thermo Scientific Forma Steri-Cycle CO 2 Incubator (Thermo Fisher Scientific, Waltham, MA, USA) [ , ].

Techniques: Lactate Dehydrogenase Assay, Infection, Incubation, Lysis, Positive Control, Comparison