Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Luteolin Inhibits the Biofilm Formation and Cytotoxicity of Methicillin-Resistant Staphylococcus aureus via Decreasing Bacterial Toxin Synthesis

doi: 10.1155/2022/4476339

Figure Lengend Snippet: Luteolin decreased the cytotoxicity of MRSA, and downregulated the expression of the hemolysin and hlaA genes in MRSA (a) Human bronchial epithelial cells were infected with luteolin-treated MRSA N315, and then the viability of human bronchial epithelial cells was measured using the CCK-8 assay. (b) Expressions of α -hemolysin and δ -hemolysin in luteolin-treated MRSA N315 were evaluated by western blot. (c) Level of hlaA in luteolin-treated MRSA N315 was detected using RT-PCR, and 16S was used as an endogenous control. ∗∗∗ P < 0.001, vs. Blank; ## P < 0.01, ### P < 0.001 vs. Control. (MRSA: methicillin-resistant Staphylococcus aureus , CCK-8: Cell Counting Kit-8, 16S: 16S rRNA, a -hemolysin: alpha hemolysin, d -hemolysin: delta hemolysin, RT-PCR: Real-Time PCR).

Article Snippet: Human bronchial epithelial cells (HBEs; CL-0346) were purchased from Procell (Wuhan, China) and grown in the specific cell medium (CM-0346, Procell, Wuhan, China).

Techniques: Expressing, Infection, CCK-8 Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control, Cell Counting, Real-time Polymerase Chain Reaction

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Long non-coding RNA NKILA inhibits proliferation and migration of lung cancer via IL-11/STAT3 signaling

doi:

Figure Lengend Snippet: The expression of NKILA was higher in normal human bronchial epithelial cell line. For (A) and (B), the expression level of lncRNA-NKILA was analyzed by Q-PCR. The expression of NKILA was analyzed in NSCLC tissues (C), Kaplan-Meier analyses of the correlations between lncRNA-NKILA expression level and survival (D). Values are means ± SEM for n = 7-8. *P < 0.05, **P < 0.01, ***P < 0.001 vs. control.

Article Snippet: Cell culture A normal human bronchial epithelial cell line (16HBE) and NSCLC adenocarcinoma cell lines (A549, NCI-H1975) were purchased from the Biopike Biological company.

Techniques: Expressing, Control

Journal: Journal of Virology

Article Title: Discovery and Evaluation of Entry Inhibitors for SARS-CoV-2 and Its Emerging Variants

doi: 10.1128/JVI.01437-21

Figure Lengend Snippet: Measurement of cytotoxicity of five drug-like compounds using MTT assay. Measurement of cytotoxicity of five drug-like compounds. (A–E) Viability of HEK293T-hACE2 cells in the presence of an indicated concentration of the compounds. (F, G) Measurement of cytotoxicity of MU-UNMC-1 and MU-UNMC-2 in Vero-STAT1 KO cells in the presence of an indicated concentration of the compounds. (H, I) Measurement of cytotoxicity of MU-UNMC-1 and MU-UNMC-2 in UNCN1T cells in the presence of an indicated concentration of the compounds. (J, K) Measurement of cytotoxicity of MU-UNMC-1 and MU-UNMC-2 in Calu-3 cells in the presence of an indicated concentration of the compounds.

Article Snippet: UNCN1T cells (a human bronchial epithelial cell line; Kerafast catalog number ENC011) were cultured in BEGM media (Bronchial Epithelial Cell Growth Medium; Lonza catalog number CC-3170) in FNC (Athena Enzyme Systems catalog number 0407) coated 96-well plates.

Techniques: MTT Assay, Concentration Assay

Journal: Journal of Virology

Article Title: Discovery and Evaluation of Entry Inhibitors for SARS-CoV-2 and Its Emerging Variants

doi: 10.1128/JVI.01437-21

Figure Lengend Snippet: SARS-CoV-2 dose-response curve in MU-UNMC-1 and MU-UNMC-2 treated and SARS-CoV-2 infected UNCN1T and Vero-STAT1 knockout cells. (A, B) MU-UNMC-1 (in blue) and MU-UNMC-2 (in green) dose-response curve by percentage inhibition of SARS-CoV-2 replication 24 and 48 hpi in UNCN1T cells with indicated drug concentrations. (C, D) MU-UNMC-1 (in blue) and MU-UNMC-2 (in green) dose-response curve by percentage inhibition of SARS-CoV-2 replication 24 and 48 hpi in Vero-STAT1 knockout cells with indicated compound concentrations.

Article Snippet: UNCN1T cells (a human bronchial epithelial cell line; Kerafast catalog number ENC011) were cultured in BEGM media (Bronchial Epithelial Cell Growth Medium; Lonza catalog number CC-3170) in FNC (Athena Enzyme Systems catalog number 0407) coated 96-well plates.

Techniques: Infection, Knock-Out, Inhibition

Journal: Journal of Virology

Article Title: Discovery and Evaluation of Entry Inhibitors for SARS-CoV-2 and Its Emerging Variants

doi: 10.1128/JVI.01437-21

Figure Lengend Snippet: Combinational effect of remdesivir and MU-UNMC-1 treatment against SARS-CoV-2 infected UNCN1T cells at 24 h postinfection. (A) Dose response curve of remdesivir in SARS-CoV-2 infected UNCN1T cells at 24 hpi in the presence of different fixed concentrations of MU-UNMC-1; (B) Dose-response curve of MU-UNMC-1 in SARS-CoV-2 infected UNCN1T cells at 24 hpi in the presence of a different fixed concentration of remdesivir; (C) Dose-response percent inhibition matrix of single and combined treatment of remdesivir and MU-UNMC-1 in SARS-CoV-2 infected UNCN1T cells at 24 hpi. (D) 3-D interaction landscape between remdesivir and MU-UNMC-2 calculated based on Loewe additive model using SynergyFinder v.2 in SARS-CoV-2 infected UNCN1T cells at 24 hpi (Loewe synergy score -30.69; with most synergistic area score of -21.34).

Article Snippet: UNCN1T cells (a human bronchial epithelial cell line; Kerafast catalog number ENC011) were cultured in BEGM media (Bronchial Epithelial Cell Growth Medium; Lonza catalog number CC-3170) in FNC (Athena Enzyme Systems catalog number 0407) coated 96-well plates.

Techniques: Infection, Concentration Assay, Inhibition

Journal: Journal of Virology

Article Title: Discovery and Evaluation of Entry Inhibitors for SARS-CoV-2 and Its Emerging Variants

doi: 10.1128/JVI.01437-21

Figure Lengend Snippet: Combinational effect of remdesivir and MU-UNMC-2 treatment against SARS-CoV-2 infected UNCN1T cells at 24 h postinfection. (A) Dose-response curve of remdesivir in SARS-CoV-2 infected UNCN1T cells at 24 hpi in the presence of a different fixed concentration of MU-UNMC-2; (B) dose-response curve of MU-UNMC-2 in SARS-CoV-2 infected UNCN1T cells at 24 hpi in the presence of a different fixed concentration of remdesivir; (C) dose-response percent inhibition matrix of single and combined treatment of remdesivir and MU-UNMC-2 in SARS-CoV-2 infected UNCN1T cells at 24 hpi. (D) 3-D interaction landscape between remdesivir and MU-UNMC-2 calculated based on Loewe additive model using SynergyFinder v.2 in SARS-CoV-2 infected UNCN1T cells at 24 hpi (Loewe synergy score 26.63; with most synergistic area score of 37.25).

Article Snippet: UNCN1T cells (a human bronchial epithelial cell line; Kerafast catalog number ENC011) were cultured in BEGM media (Bronchial Epithelial Cell Growth Medium; Lonza catalog number CC-3170) in FNC (Athena Enzyme Systems catalog number 0407) coated 96-well plates.

Techniques: Infection, Concentration Assay, Inhibition

Journal: Frontiers in Microbiology

Article Title: The Pseudomonas aeruginosa Secreted Protein PA3611 Promotes Bronchial Epithelial Cell Epithelial-Mesenchymal Transition via Integrin αvβ6-Mediated TGF-β1-Induced p38/NF-κB Pathway Activation

doi: 10.3389/fmicb.2021.763749

Figure Lengend Snippet: Effects of PA3611 on the proliferation and epithelial-mesenchymal transition (EMT) of bronchial epithelial cells. 16HBE14o- and RTE cells were treated with different amounts of PA3611 (1, 5, 10, and 50 μg) for 24, 48, or 72 h. A CCK-8 assay was performed to estimate cell proliferation (A,B) . Transmission electron microscopy was used to observe the ultrastructure of the above cells treated with PA3611 (10 μg/mL) for 48 h (C) . EMT-related protein (α-SMA, vimentin, E -cadherin and ZO-1) levels in 16HBE14o- and RTE cells at 48 h were assessed by Western blot analysis (D–F) . Fluorescence intensities of EMT-related proteins (α-SMA, vimentin, E -cadherin and ZO-1) were assessed by immunofluorescence staining in 16HBE14o- and RTE cells at 48 h (F–H) , and photographs were obtained with a confocal laser microscope (100 × objective lens). All the presented graphs are representative of the results of three independent experiments. The data are presented as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. the control group. White arrow ① and ② nucleus; ③ cells crest; ④ cells vacuolation; ⑤ intercellular gaps; ⑥ intracellular glycogen; ⑦ mitochondria.

Article Snippet: The human bronchial epithelial cell line 16HBE14o-, which was originally created by Prof. D. Gruenert , and the rat bronchial epithelial cell line RTE were purchased from Procell Company (Wuhan, China) and preserved in our laboratory.

Techniques: CCK-8 Assay, Transmission Assay, Electron Microscopy, Western Blot, Fluorescence, Immunofluorescence, Staining, Microscopy, Control

Journal: Frontiers in Microbiology

Article Title: The Pseudomonas aeruginosa Secreted Protein PA3611 Promotes Bronchial Epithelial Cell Epithelial-Mesenchymal Transition via Integrin αvβ6-Mediated TGF-β1-Induced p38/NF-κB Pathway Activation

doi: 10.3389/fmicb.2021.763749

Figure Lengend Snippet: Effects of PA3611 and TGF-β1 on the expression of p65 phosphorylation and EMT-related markers. 16HBE14o- and RTE cells were treated with PA3611 (10 μg/mL), and the TGF-β1 levels in 16HBE14o- and RTE cell culture supernatants were detected by an enzyme-linked immunosorbent assay (ELISA) at 24, 48, and 72 h (A,B) . 16HBE14o- and RTE cells were treated with PA3611 (10 μg), TGF-β1 (2 ng/mL) or PA3611 (10 μg) plus TGF-β1-neutralizing antibody (10 μg/mL) for 48 h, and EMT-related markers (α-SMA, vimentin, E -cadherin and ZO-1, C–G ) and p65 phosphorylation (H,I) were then assessed by Western blot analysis. All the presented graphs are representative of the results of three independent experiments. The data are presented as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. the control group.

Article Snippet: The human bronchial epithelial cell line 16HBE14o-, which was originally created by Prof. D. Gruenert , and the rat bronchial epithelial cell line RTE were purchased from Procell Company (Wuhan, China) and preserved in our laboratory.

Techniques: Expressing, Phospho-proteomics, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Frontiers in Microbiology

Article Title: The Pseudomonas aeruginosa Secreted Protein PA3611 Promotes Bronchial Epithelial Cell Epithelial-Mesenchymal Transition via Integrin αvβ6-Mediated TGF-β1-Induced p38/NF-κB Pathway Activation

doi: 10.3389/fmicb.2021.763749

Figure Lengend Snippet: Effects of p65 overexpression or knockdown on the expression of TGF-β1, p38 phosphorylation and the expression of EMT-related markers. 16HBE14o- and RTE cells were transfected with a control vector (indicated as blank), a p65 overexpression vector (indicated as p65), or a specific siRNA directed against p65 (indicated as sh-p65). Twenty-four hours post-transfection, cells were treated with PBS (negative control) or PA3611 (10 μg/mL) for another 48 h. Then, cells were collected to examine the gene and protein expression of p38, p65 and EMT-related markers (A–G,I–N) . Cell culture supernatants were collected for TGF-β1 level evaluation by enzyme-linked immunosorbent assay (H) . The results are representative of three independent experiments. The data are presented as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. the control group.

Article Snippet: The human bronchial epithelial cell line 16HBE14o-, which was originally created by Prof. D. Gruenert , and the rat bronchial epithelial cell line RTE were purchased from Procell Company (Wuhan, China) and preserved in our laboratory.

Techniques: Over Expression, Knockdown, Expressing, Phospho-proteomics, Transfection, Control, Plasmid Preparation, Negative Control, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Microbiology

Article Title: The Pseudomonas aeruginosa Secreted Protein PA3611 Promotes Bronchial Epithelial Cell Epithelial-Mesenchymal Transition via Integrin αvβ6-Mediated TGF-β1-Induced p38/NF-κB Pathway Activation

doi: 10.3389/fmicb.2021.763749

Figure Lengend Snippet: PA3611 but not TGF-β1 stimulated avβ6 protein expression. 16HBE14o- and RTE cells were treated with different concentrations of PA3611 (1, 5, and 10 μg/mL) for different durations (12, 24, and 48 h), and αvβ6 expression was detected by Western blotting (A–H) . 16HBE14o- and RTE cells were treated with PBS, TGF-β1 (2 ng/mL), PA3611 (10 μg/mL), and PA3611 (10 μg/mL) + TGF-β1-neutralizing antibody (10 μg/mL) for 48 h, and αvβ6 expression was detected by immunofluorescence staining (I,J) and Western blot (K,L) . The results are representative of three independent experiments. The data are presented as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. the control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. the TGF-β1 group.

Article Snippet: The human bronchial epithelial cell line 16HBE14o-, which was originally created by Prof. D. Gruenert , and the rat bronchial epithelial cell line RTE were purchased from Procell Company (Wuhan, China) and preserved in our laboratory.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Control

Journal: Frontiers in Microbiology

Article Title: The Pseudomonas aeruginosa Secreted Protein PA3611 Promotes Bronchial Epithelial Cell Epithelial-Mesenchymal Transition via Integrin αvβ6-Mediated TGF-β1-Induced p38/NF-κB Pathway Activation

doi: 10.3389/fmicb.2021.763749

Figure Lengend Snippet: PA3611 stimulated TGF-β1 expression and induced EMT, which could be inhibited by blocking integrin αvβ6. 16HBE14o- cells were pretreated with the integrin αvβ6-blocking antibody 10D5 (10 μM) or a specific inhibitor of TGF-β1-Smad2/3 signaling, SB431542 (10 μM), for 2 h prior to incubation with PA3611 (10 μg/mL) for 48 h. The TGF-β1 levels in cell culture supernatants were evaluated by ELISA (A) . EMT-related proteins (B–F) and signaling pathway proteins (G–I) were detected using Western blotting. The results are representative of three independent experiments, and the data are presented as the means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. the control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. the PA3611 group.

Article Snippet: The human bronchial epithelial cell line 16HBE14o-, which was originally created by Prof. D. Gruenert , and the rat bronchial epithelial cell line RTE were purchased from Procell Company (Wuhan, China) and preserved in our laboratory.

Techniques: Expressing, Blocking Assay, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Frontiers in Microbiology

Article Title: The Pseudomonas aeruginosa Secreted Protein PA3611 Promotes Bronchial Epithelial Cell Epithelial-Mesenchymal Transition via Integrin αvβ6-Mediated TGF-β1-Induced p38/NF-κB Pathway Activation

doi: 10.3389/fmicb.2021.763749

Figure Lengend Snippet: Effects of p38 overexpression or knockdown on the expression of TGF-β1, EMT-related marker expression and the phosphorylation of p38 and p65. 16HBE14o- cells were transfected with a control vector (indicated as blank), a p38 overexpression vector (indicated as p38), or a specific siRNA directed against p38 (indicated as sh-p38). Twenty-four hours post-transfection, cells were treated with PBS (negative control) or PA3611 (10 μg/mL) for another 48 h. Then, cell culture supernatants were collected for TGF-β1 level evaluation by enzyme-linked immunosorbent assay (H) , and cells were collected to examine the gene and protein expression of p38, p65 and EMT-related markers (A–G,I–N) . The results are representative of three independent experiments. The data are presented as the means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, vs. the control group.

Article Snippet: The human bronchial epithelial cell line 16HBE14o-, which was originally created by Prof. D. Gruenert , and the rat bronchial epithelial cell line RTE were purchased from Procell Company (Wuhan, China) and preserved in our laboratory.

Techniques: Over Expression, Knockdown, Expressing, Marker, Phospho-proteomics, Transfection, Control, Plasmid Preparation, Negative Control, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Clinical and Experimental Pathology

Article Title: CXCR4 inhibitor attenuates allergen-induced lung inflammation by down-regulating MMP-9 and ERK1/2

doi:

Figure Lengend Snippet: Results for immunostaining of CXCR4 in bronchial epithelial cells. CXCR4 in 16HBE cells were first probed with a rabbit derived mAb and then stained a green fluorescent labeled anti-rabbit IgG (green). Nuclei were stained in red by PI (original magnification × 400).

Article Snippet: Cell culture and treatment The human bronchial epithelial cell line (16HBE) was obtained from Fuxiang Biotechnology Co. Ltd (Shanghai, China).

Techniques: Immunostaining, Derivative Assay, Staining, Labeling

Journal: International Journal of Clinical and Experimental Pathology

Article Title: CXCR4 inhibitor attenuates allergen-induced lung inflammation by down-regulating MMP-9 and ERK1/2

doi:

Figure Lengend Snippet: CXCL12/CXCR4 synergizes with IL-13 to enhance epithelial MMP-9 expression. A. CXCL12 time-dependently induced epithelial cells expression of MMP-9. 16HBE cells were cultured in serum-free medium at 37°C for 24 h and then stimulated with CXCL12 (200 ng/ml) as indicated times. B. Gelatin zymographic results for conditioned media collected from CXCL12 treated 16HBE cells. C. Western blot analysis of epithelial MMP-9 expression after CXCL12 and/or IL-13 stimulation. 16HBE cells were treated for 24 h with 10 ng/ml IL-13, 200 ng/ml CXCL12, 10 ng/ml IL-13 and 200 ng/ml CXCL12, CXCL12 and saline, CXCL12 and AMD3100, respectively. The cells were then harvested for Western blot analysis of MMP-9 expression. D. A bar graphic figure showing the results of 5 independent experiments conducted. *, P < 0.05 as compared with Control group; #, P < 0.05 as compared with CXCL12 group.

Article Snippet: Cell culture and treatment The human bronchial epithelial cell line (16HBE) was obtained from Fuxiang Biotechnology Co. Ltd (Shanghai, China).

Techniques: Expressing, Cell Culture, Western Blot, Saline, Control

Journal: International Journal of Clinical and Experimental Pathology

Article Title: CXCR4 inhibitor attenuates allergen-induced lung inflammation by down-regulating MMP-9 and ERK1/2

doi:

Figure Lengend Snippet: CXCL12/CXCR4 signaling induces ERK1/2 expression and activation. 16HBE cells were incubated with 200 ng/ml CXCL12 as the indicated time and then subjected to Western blot analysis of ERK1/2 expression and activation. A. CXCL12 potently increased total ERK1/2 protein levels and pERK1/2 levels. The results are a representative of 3 independent experiments conducted. B. Blockade of ERK1/2 signaling by PD98059 abolished the stimulatory effect of CXCL12 on epithelial MMP-9 expression. 16HBE cells were pretreated with PD98059 (50 μM) for 30 min before CXCL12 stimulation. Addition of PD98059 completely diminished CXCL12 induced MMP-9 expression in 16HBE cells.

Article Snippet: Cell culture and treatment The human bronchial epithelial cell line (16HBE) was obtained from Fuxiang Biotechnology Co. Ltd (Shanghai, China).

Techniques: Expressing, Activation Assay, Incubation, Western Blot

Journal: International Journal of Molecular and Cellular Medicine

Article Title: Long Non-coding RNA snaR Promotes Proliferation in EGFR Wild Type Non-Small Cell Lung Cancer Cells

doi: 10.22088/IJMCM.BUMS.10.4.258

Figure Lengend Snippet: snaR expression in normal human bronchial epithelial cells 16HBE and NSCLC cell lines. Data are presented as mean ± SD (*P < 0.05, **P < 0.01)

Article Snippet: C ell lines and culture conditions NSCLC cell lines including two carrying wild type EGFR (A549, SPC-A1) and two carrying mutations in EGFR (PC9 and H1975: PC9 cells carry a Glu746-Ala750 deletion mutation in exon 19 of the EGFR gene, and H1975 cells carry two-point mutations, T790M and L858R, in exons 20 and 21, respectively), and a normal human bronchial epithelial cell line (16HBE) were purchased from the Cell Bank of the Pasteur Institute of Iran (Tehran, Iran).

Techniques: Expressing