Journal: Cancer Medicine

Article Title: PIAS3 expression in squamous cell lung cancer is low and predicts overall survival

doi: 10.1002/cam4.372

Figure Lengend Snippet: PIAS3 expression is low in the majority of squamous cell carcinoma lung tumors by western blotting. Protein precipitates of squamous cell lung cancer tumors from LCBRN were solubilized and analyzed for PIAS3 and β -actin expression by western blotting (top). The PIAS3 and corresponding β -actin bands were quantified using image J software and a ratio of the resulting values is shown (bottom). NL-20 and A549 protein lysates were used as controls.

Article Snippet: Human pulmonary benign and malignant epithelial cell lines, NL-20 and A549 respectively, as well as human squamous cell lung cancer cell lines Calu-1, H520, H1869, H1385, and SW900 were purchased from American Type Culture Collection (Manassas, VA) and maintained in Dulbecco's Modified Eagle Medium (DMEM)/Ham's F-12 medium supplemented with 10% (v/v) fetal bovine serum (Hyclone; ThermoFisher Scientific, Waltham, MA) and 2 mmol/L l -Glutamine (GlutaMAX; Invitrogen, Camirillo, CA) in a 5% CO 2 humidified incubator at 37°C.

Techniques: Expressing, Western Blot, Software

Journal: Cancer Medicine

Article Title: PIAS3 expression in squamous cell lung cancer is low and predicts overall survival

doi: 10.1002/cam4.372

Figure Lengend Snippet: PIAS3 expression is low in most squamous cell carcinoma cell lines by western blotting. Protein precipitates of squamous cell lung cancer cell lines were prepared and analyzed for PIAS3 and β -actin expression by western blotting (top). The PIAS3 and corresponding β -actin bands were quantified using image J software and a ratio of the resulting values is shown (bottom). NL-20 and A549 protein lysates were used as controls.

Article Snippet: Human pulmonary benign and malignant epithelial cell lines, NL-20 and A549 respectively, as well as human squamous cell lung cancer cell lines Calu-1, H520, H1869, H1385, and SW900 were purchased from American Type Culture Collection (Manassas, VA) and maintained in Dulbecco's Modified Eagle Medium (DMEM)/Ham's F-12 medium supplemented with 10% (v/v) fetal bovine serum (Hyclone; ThermoFisher Scientific, Waltham, MA) and 2 mmol/L l -Glutamine (GlutaMAX; Invitrogen, Camirillo, CA) in a 5% CO 2 humidified incubator at 37°C.

Techniques: Expressing, Western Blot, Software

Journal: Tobacco Induced Diseases

Article Title: Effect of cigarette smoke condensate on gene promoter methylation in human lung cells

doi: 10.1186/1617-9625-12-15

Figure Lengend Snippet: Heatmap of the intensity of promoter methylation of the 24-genes panel in PSAE (human primary small airway epithelial) cells. Cells were cultured in the absence or presence of 0.3 or 1.0 μg/ml CSC for 3, 14 or 30 days. Methylation was assessed by the Methyl-Profiler DNA Methylation PCR System. Methylation analysis in the lung cancer lines, A549 and H1299, is also represented. Color code and intensity indicate level of methylation.

Article Snippet: The human lung cell lines, PSAE (primary small airway epithelial) cells (PCS-301-0100), NL-20 (immortalized bronchial epithelial) cells, A549 (adenocarcinomic alveolar basal epithelial) cells and H1299 (non-small lung carcinoma) cells, were obtained from the American Type Culture Collection (ATCC) (Manassa, VA).

Techniques: Methylation, Cell Culture, DNA Methylation Assay

Journal: Tobacco Induced Diseases

Article Title: Effect of cigarette smoke condensate on gene promoter methylation in human lung cells

doi: 10.1186/1617-9625-12-15

Figure Lengend Snippet: Heatmap of the intensity of promoter methylation of the 24-genes panel in NL-20 (human immortalized bronchial epithelial) cells. Cells were cultured in the absence or presence of 10 or 100 μg/ml CSC for 30 days. Methylation was assessed by the Methyl-Profiler DNA Methylation PCR System. Methylation analysis in the lung cancer lines, A549 and H1299, is also represented. Color code and intensity indicate level of methylation.

Article Snippet: The human lung cell lines, PSAE (primary small airway epithelial) cells (PCS-301-0100), NL-20 (immortalized bronchial epithelial) cells, A549 (adenocarcinomic alveolar basal epithelial) cells and H1299 (non-small lung carcinoma) cells, were obtained from the American Type Culture Collection (ATCC) (Manassa, VA).

Techniques: Methylation, Cell Culture, DNA Methylation Assay

Journal: Communications Biology

Article Title: H5N1 infection impairs the alveolar epithelial barrier through intercellular junction proteins via Itch-mediated proteasomal degradation

doi: 10.1038/s42003-022-03131-3

Figure Lengend Snippet: A549 ( a ), NL20 ( b ), and MDCK ( c ) cells were infected with various MOI of H5N1 viruses (the SY strain) for 24 h or with H5N1 viruses (1 MOI) for the indicated lengths of time. Cell lysates were analyzed for the levels of Gli1, Snail, and four junction proteins by Western blot. The density of the bands was analyzed by using NIH Image-J software and normalized by the arbitrary units of β-actin. Data are the mean ± SD of three experiments. *p < 0.05, **p < 0.01, com p ared to uninfected control. d , e The effect of influenza viruses on GLI1 , SNAI , CDH , CLDN1, and OCLN mRNA expression. Total mRNA from A549 cells infected with the indicated MOI of H5N1 ( d ) or H1N1 ( e ) viruses was analyzed for GLI1 , SNAI , OCLN, CLDN1, and CDH mRNA levels by RT-PCR. Data are the mean ± SD of three independent experiments. **p < 0.01, compared to uninfected control. * p < 0.05, ** p < 0.01.

Article Snippet: A549 (a human lung cancer cell line of alveolar epithelial cell origin) (CCR-185), NL20 (a human non-tumoral alveolar epithelial cell line) (CRL-1503), and MDCK (Madin-Darby canine kidney) (CCR-34) cells were purchased from the American Tissue Culture Collection (Manassas, VA).

Techniques: Infection, Western Blot, Software, Control, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Communications Biology

Article Title: H5N1 infection impairs the alveolar epithelial barrier through intercellular junction proteins via Itch-mediated proteasomal degradation

doi: 10.1038/s42003-022-03131-3

Figure Lengend Snippet: a H5N1 viruses induce TAK1 phosphorylation. A549, MDCK, and NL20 cells were infected with various MOI of the H5N1 viruses for 24 h or with the H5N1 viruses (1 MOI) for the indicated lengths of time. Cell lysates were analyzed for the levels of TAK1 phosphorylation, NP, NS1, and β-actin by Western blot. b 5Z blocks H5N1 virus-induced Itch expression and junction protein downregulation. A549 cells were left uninfected or infected with 1 MOI of H5N1 viruses. After incubation for 12 h, 5Z (5 μM) was added and then incubated for another 12 h. Cell lysates were analyzed by Western blot for TAK1, ERK, JNK, p38, and p65 phosphorylation and for the levels of junction proteins with their specific antibodies. The density of bands was analyzed using NIH Image-J software and normalized by the arbitrary units of β-actin. Data are the mean ± SD of three experiments. *p < 0.05, **p < 0.01. c 5Z blocks H5N1 virus-induced disruption of intercellular junction structure. A549 cells seeded on coverslips were treated as above. Cells were immunostained with antibodies against occludin and claudin-1 and visualized under a confocal microscope. d Quantification of fluorescence signals. The monolayers immunostained for occludin and claudin-1 in c were analyzed for immunofluorescence intensity by using Image J software. The fluorescent signals of intercellular junctions were quantified and plotted as bar graphs. Data represent the mean ± SD of five random fields (40X) from one of three independent experiments with similar results. e 5Z blocks the H5N1 virus-induced decrease of electronic resistance. A549 cells seeded in Transwell inserts were left uninfected or infected with H5N1 viruses. After incubation for 12 h, 5Z (5 μM) was added and then incubated for another 12 h. TEER was measured at the indicated times. The results represent the mean ± SD of three independent experiments. ** p < 0.01.

Article Snippet: A549 (a human lung cancer cell line of alveolar epithelial cell origin) (CCR-185), NL20 (a human non-tumoral alveolar epithelial cell line) (CRL-1503), and MDCK (Madin-Darby canine kidney) (CCR-34) cells were purchased from the American Tissue Culture Collection (Manassas, VA).

Techniques: Phospho-proteomics, Infection, Western Blot, Virus, Expressing, Incubation, Software, Disruption, Microscopy, Fluorescence, Immunofluorescence