Journal: Cell reports

Article Title: Interferon (IFN)-γ promotes monocyte-mediated lung injury during influenza infection

doi: 10.1016/j.celrep.2022.110456

Figure Lengend Snippet: Key Resources Table

Article Snippet: Primary Bronchial/Tracheal Epithelial Cells; Normal, Human , ATCC , Cat#PCS-300-01.

Techniques: Virus, Recombinant, cDNA Synthesis, SYBR Green Assay, Protein Extraction, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Software, Microscopy, Flow Cytometry, Fluorescence

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 1: ENDS aerosol exposure causes increased cell death in human lung epithelial cells The BEAS-2B cell line was exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N-), 100μM Nicotine (Nicotine), HEPA filtered air (Air), and room temperature (Untreated) for 4 min. Cell death was analyzed 24h after treatment by flow cytometry using Caspase-3/7 stain and SYTOXTM permeability dye. Representative plots are shown in (A) and quantification show in (B-E). Necrotic cells are defined as SYTOXTM +, Caspase-3/7- (i) (B). Secondary necrotic cells are dual positive (ii) (C). Apoptotic cells are Caspase-3/7+, SYTOXTM- (iii) (D). All cells with activated Caspase-3/7 (E). Experiments were performed 3 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, &=p<0.05 compared to PG/VG N-, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol, Flow Cytometry, Staining, Permeability

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 2: ENDS aerosol exposure causes increased cell death in mouse lung epithelial cells The MLE12 cell line was exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N-), 100μM Nicotine (Nicotine), HEPA filtered air (Air), and room temperature (Untreated) for 4 min. Cell death was analyzed 24h after treatment by flow cytometry using Caspase-3/7 stain and SYTOXTM permeability dye. Representative plots are shown in (A) and quantification show in (B-E). Necrotic cells are defined as SYTOXTM +, Caspase-3/7- (i) (B). Secondary necrotic cells are dual positive (ii) (C). Apoptotic cells are Caspase-3/7+, SYTOXTM- (iii) (D). All cells with activated Caspase-3/7 (E). Experiments were performed 3 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, &=p<0.05 compared to PG/VG N-, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol, Flow Cytometry, Staining, Permeability

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 3: One week of ENDS aerosol exposure causes decreased trans-epithelial resistance, dissociation of cells, and cell death Primary human bronchial epithelial cells were differentiated on transwell inserts into a pseudostratified epithelial layer. These cells were exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N- ), HEPA filtered air (Air), 100uM nicotine (Nicotine), and room temperature (Untreated) for 4 min. Trans- epithelial resistance was measured using a WPI EVOM TEER meter and STX3 electrode. Measurements were taken in triplicate and averaged. Background readings of empty wells as well as baseline measurements of each sample were used to calculate the change in resistance over the course of the treatment. Dissociated and dead cells were detected using trypan blue exclusion and counting on a hemocytometer. Treatments occurred once per day for 7 days. Experiments were performed 2 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol

Journal: Purinergic Signalling

Article Title: Autocrine signaling via release of ATP and activation of P2X7 receptor influences motile activity of human lung cancer cells

doi: 10.1007/s11302-014-9411-x

Figure Lengend Snippet: TGF-β1-induced migration is mediated by release of ATP and activation of P2X7 receptor in H292 lung cancer cells, but not BEAS-2B normal lung cells. a The expression of P2X7 receptor in BEAS-2B, A549, PC-9, and H292 cells was detected by immunoblotting as described in “Materials and methods”. b, c BEAS-2B cells (b) or H292 cells (c) were stimulated with TGF-β1 (5 ng/mL) and incubated for the indicated times, then the concentration of ATP in the culture medium was measured as described in “Materials and methods”. d, e Cell migration was examined by means of Transwell assay as described in “Materials and methods”. BEAS-2B cells (d) or H292 cells (e) were pretreated for 30 min with apyrase (20 U/mL) or A438079 (100 μM) and then stimulated for 24 h with TGF-β1 (5 ng/mL). The lower membrane surfaces were photographed through a microscope at ×20 magnification, and migrated cells in each field were counted. Values are means ± SE (n = 4–10). A significant difference between the indicated group and control group is indicated by *** or ### (P < 0.001), ** (P < 0.01), †(P < 0.05)

Article Snippet: Cell culture BEAS-2B normal human bronchial epithelial cell line and NCI-H292 human lung mucoepidermoid carcinoma cell line were purchased from DS Pharma Biomedical (Osaka, Japan).

Techniques: Migration, Activation Assay, Expressing, Western Blot, Incubation, Concentration Assay, Transwell Assay, Microscopy

Journal: Molecular Therapy

Article Title: Long Non-coding RNA BGas Regulates the Cystic Fibrosis Transmembrane Conductance Regulator

doi: 10.1038/mt.2016.112

Figure Lengend Snippet: BGas and as4 mediated regulation of CFTR . ( a ) A schematic depicting the CFTR locus with transcriptional start sites (TSS) for CFTR , EST BG213071 (BGas), and those primers used to evaluate CFTR expression. The sasRNA target site (as4) in the BGas promoter is also shown along with the two exons making up BGas. ( b ) The effects of BGas over-expression using an exogenously expressed BGas (exBGas) on CFTR expression in 1HAEo- cells. The control (pcDNA3.1) and exBGas transfected cells are shown. ( c,d ) The effects of sasRNA as4 relative to control (pU6M2) on ( c ) BGas and ( d ) CFTR expression in 1HAEo- cells. ( e,f ) The effects of exBGas ( e ) and sasRNA as4 ( f ) on CFTR expression in CFPAC cells. ( g ) Dose-dependent effect of an siRNA targeting the as4 site (siRNA4) on ΔF508- CFTR chloride channel transport function in CF Human Bronchial Epithelial (CFhBE) primary cells (epithelial voltage clamp assay, Ussing chamber). ( h ) The localization of biotin containing BGas at the intergenic locus of CFTR in transfected CFPAC cells contrasted with the lambda biotin control. For b–f , the averages of triplicate treated cultures are shown with the standard error of the means and a P value from a paired T -test, * P < 0.05 and ** P < 0.01.

Article Snippet: Briefly, primary human bronchial epithelial (hBE) cells from a CF patient homozygous for ΔF508- CFTR (Asterand Biosciences, Royston, UK) were plated onto Costar Snapwell (Corning, Corning, NY) tissue-culture inserts coated with collagen type IV, and cultured in Bronchial epithelial growth medium (Lonza, Basel, Switzerland).

Techniques: Expressing, Over Expression, Transfection

Journal: Neoplasia (New York, N.Y.)

Article Title: Multimutated Herpes Simplex Virus G207 Is a Potent Inhibitor of Angiogenesis 1

doi:

Figure Lengend Snippet: Susceptability of human endothelial cells (HUVEC and HDMEC), fibroblasts (HFF), retinal glial cells (HRG), normal human bronchial epithelial cells (NHBE), and alveolar rhabdomyosarcoma (KFR) cell cultures to G207 and HSV-1 wild-type strain McIntyre. Cells were infected with G207 or McIntyre at an MOI of 0.1. Each data point (mean of triplicate wells ± SD) is the percentage of surviving cells compared with mock-infected cells in control wells or infectious viral titers at each time point, respectively. Some error bars for data points are smaller than the symbol size.

Article Snippet: Normal human bronchial epithelial cells (NHBE) were obtained from Clonetics (CellSystems, St. Katharinen, Germany) and cultured in BEGM medium according to the instructions of the producer.

Techniques: Infection, Control