Journal: Mediators of Inflammation

Article Title: Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

doi: 10.1155/2014/136463

Figure Lengend Snippet: Protein expression of HVEM, LT β R, and BTLA on purified human basophils, purified human eosinophils, and human bronchial epithelial BEAS-2B cells. Representative histograms of the cell surface expression of HVEM, LT β R, and BTLA on (a, b, c) basophils (d, e, f), eosinophils, and (g, h, i) BEAS-2B cells determined with gating by respective side scatter and forward scatter using flow cytometry were obtained from triplicate experiments with essentially identical results.

Article Snippet: The human bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Purification, Flow Cytometry

Journal: Mediators of Inflammation

Article Title: Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

doi: 10.1155/2014/136463

Figure Lengend Snippet: Effect of LIGHT on the cell surface expression of ICAM-1 on BEAS-2B cells or basophils/eosinophils. Expressions of ICAM-1 on BEAS-2B cells alone or in the coculture, and basophils/eosinophils in the coculture with or without LIGHT stimulation are presented with representative bar charts. (a) Representative histogram of the cell surface expression of ICAM-1 on BEAS-2B cells (1 × 10 5 cells) treated with different concentration of LIGHT (0–100 ng/mL) for 24 h is shown. (b) Basophils or (c) eosinophils (3 × 10 5 cells) and confluent BEAS-2B cells (1 × 10 5 cells) were cultured either together or separately with or without LIGHT (1–100 ng/mL) for 24 h. Surface expressions of ICAM-1 on 5,000 BEAS-2B cells, basophils, or eosinophils are expressed as the mean plus SEM of MFI of three independent experiments with three blood samples. * P < 0.05, *** P < 0.001. (d) Basophils or (e) eosinophils (0.3 × 10 5 –3 × 10 5 cells) and confluent BEAS-2B cells (1 × 10 5 cells) were cultured together with or without LIGHT (100 ng/mL) and transwell (pore size 0.4 μ m) for 24 h. Surface expressions of ICAM-1 on 5,000 BEAS-2B cells, basophils, or eosinophils are expressed as the mean plus SEM of MFI of three independent experiments with three blood samples. * P < 0.05; ** P < 0.01 when compared with the cocultured BEAS-2B cells and eosinophils without stimulation with LIGHT (empty bar). # P < 0.05 when compared with the corresponding group without transwell inserts. BS: BEAS-2B cells alone; CoBS: BEAS-2B cells in the coculture; CoBAS: basophils in the coculture; CoEosi: eosinophils in the coculture.

Article Snippet: The human bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Concentration Assay, Cell Culture

Journal: Mediators of Inflammation

Article Title: Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

doi: 10.1155/2014/136463

Figure Lengend Snippet: Adhesion of basophils or eosinophils onto BEAS-2B cells. Coculture of BEAS-2B cells (1 × 10 5 cells) and basophils/eosinophils (3 × 10 5 cells) using transwell inserts or BEAS-2B cells alone were stimulated by LIGHT (0–100 ng/mL) for 24 h. Basophils/eosinophils were removed and freshly isolated basophils/eosinophils (3 × 10 5 cells) were added into the corresponding well containing the adherent BEAS-2B cells (1 × 10 5 cells) for 1 h incubation. The ratio of (a) basophils or (b) eosinophils adherent onto BEAS-2B was analysed using flow cytometry as described in . Results are expressed as the mean plus SEM of three independent experiments with three blood samples. * P < 0.05, *** P < 0.001. BEAS-2B: BEAS-2B cells alone were treated with LIGHT prior to adhesion analysis; Co-BAS + BEAS-2B: Coculture of BEAS-2B cells and basophils were treated with LIGHT before BEAS-2B cells were used for adhesion analysis; Co-Eosi + BEAS-2B: Coculture of BEAS-2B cells and eosinophils were treated with LIGHT before BEAS-2B cells were used for adhesion analysis. In the above adhesion assay, basophils/eosinophils and BEAS-2B cells were analyzed separately using flow cytometry. (c) Representative histograms of the flow cytometric analysis of the number of adherent basophils in coculture gated by the specific cell surface basophilic marker CD203c were shown. (d) Representative dot plots of the flow cytometric analysis of the number of adherent eosinophils in coculture gated by the SSC and FSC were shown.

Article Snippet: The human bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Isolation, Incubation, Flow Cytometry, Cell Adhesion Assay, Marker

Journal: Mediators of Inflammation

Article Title: Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

doi: 10.1155/2014/136463

Figure Lengend Snippet: Effects of LIGHT on the release of IL-6 and CXCL8 from the coculture of basophils/eosinophils and BEAS-2B cells. Confluent BEAS-2B cells (1 × 10 5 cells) and (a, b) basophils/(c, d) eosinophils (3 × 10 5 cells) were cultured either together or separately with or without LIGHT for 24 h. Release of IL-6 and CXCL8 in culture supernatants was determined by Milliplex human cytokine/chemokine magnetic panel assay. Results are expressed as the mean plus SEM of three independent experiments with three blood samples. * P < 0.05, ** P < 0.01, *** P < 0.001. BAS: basophils; Eosi: eosinophils; BS: BEAS-2B cells; CO: coculture.

Article Snippet: The human bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Cell Culture

Journal: Mediators of Inflammation

Article Title: Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

doi: 10.1155/2014/136463

Figure Lengend Snippet: Effects of LIGHT on the release of MMP-9 in the coculture of basophils and BEAS-2B cells. Confluent BEAS-2B cells (1 × 10 5 cells) and basophils (3 × 10 5 cells) were cultured either together or separately with LIGHT (0–100 ng/mL) for 24 h. Release of MMP-9 in culture supernatants was measured by Milliplex human MMP panel assay. Results are expressed as the mean plus SEM of three independent experiments with three blood samples. * P < 0.05, *** P < 0.001. BAS: basophils; BS: BEAS-2B cells; CO: coculture of basophils and BEAS-2B cells.

Article Snippet: The human bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Cell Culture

Journal: Mediators of Inflammation

Article Title: Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

doi: 10.1155/2014/136463

Figure Lengend Snippet: Phosphorylation of p38 MAPK, ERK1/2, and I κ B α in BEAS-2B cells upon the coculture of basophils/eosinophils and BEAS-2B cells with the stimulation of LIGHT. BEAS-2B cells (1 × 10 5 cells) and basophils/eosinophils (3 × 10 5 cells) were cultured either together or separately with or without LIGHT stimulation (100 ng/mL) for different time points (0, 15, 30, and 60 min). The intracellular expression of phosphorylated (p) p38 MAPK, pERK1/2, and pI κ B α in (a) permeabilized basophils and (b) eosinophils alone without coculture, and (c) pp38 MAPK, (d) pERK1/2, and (e) pI κ B α of permeabilized BEAS-2B cells in with or without coculture with basophils/eosinophils were measured by intracellular immunofluorescence staining using flow cytometry. Results are shown in MFI and expressed as the arithmetic mean plus SEM of three independent experiments with three blood samples in bar charts. * P < 0.05, ** P < 0.01 when compared with coculture control group. BAS: basophils; Eosi: eosinophils; BS: BEAS-2B cells.

Article Snippet: The human bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Cell Culture, Expressing, Immunofluorescence, Staining, Flow Cytometry

Journal: Mediators of Inflammation

Article Title: Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

doi: 10.1155/2014/136463

Figure Lengend Snippet: Effects of signaling molecule inhibitors on the cell surface expression of ICAM-1 and induction of cytokines/chemokines from coculture of BEAS-2B cells and basophils/eosinophils in the presence of LIGHT (100 ng/mL). Cocultures of (a, b, c, d) basophils or (e, f) eosinophils (3 × 10 5 cells) and confluent BEAS-2B cells (1 × 10 5 cells) were pretreated with BAY11-7082 (1 μ M), SP600125 (3 μ M), SB203580 (7.5 μ M), U0126 (10 μ M), or LY294002 (10 μ M) for 1 h, followed by incubation with or without LIGHT (100 μ g/mL) in the presence of inhibitors for a further 24 h. Cell surface expression of ICAM-1 on 5,000 cells was analyzed by flow cytometry as MFI. Results are expressed as mean plus SEM of three independent experiments with three blood samples in bar charts. Release of cytokines/chemokines and MMP-9 in culture supernatant was measured by Milliplex assay. Results are expressed as mean plus SEM. DMSO (0.1%) was used as the vehicle control. * P < 0.05, ** P < 0.01 when compared with vehicle or negative control.

Article Snippet: The human bronchial epithelial cell line (BEAS-2B) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Incubation, Flow Cytometry, Negative Control

Journal: Bioscience Reports

Article Title: Metabolic profiling and combined therapeutic strategies unveil the cytotoxic potential of selenium-chrysin (SeChry) in NSCLC cells

doi: 10.1042/BSR20240752

Figure Lengend Snippet: ( A ) SeChry@PURE G4 -LA 24 nanoformulation. Chemical structures of the generation four polyurea dendrimer lactate conjugate and encapsulated SeChry molecules. A549, H292, PC-9, H1975, H522, H596 and BEAS-2B cells were exposed to empty nanoparticles (PURE G4 -LA) or SeChry@PURE G4 -LA 24 for 48 h. Percentage of cell death analyzed by flow cytometry using Annexin V and PI in ( B ) PURE G4 -LA 24 -treated cells and ( C ) Sechry@PURE G4 -LA 24 -treated cells. ( D ) EC50 curves and values for SeChry@PURE G4 -LA. Data are represented as mean ± SD. * P <0.5, ** P <0.01, *** P <0.001, **** P <0.0001 (one-way ANOVA with Tukey’s multiple comparisons test was used).

Article Snippet: Normal human bronchial epithelium cell line BEAS-2B (CRL-3588™, ATCC) was also used.

Techniques: Flow Cytometry

Journal: Bioscience Reports

Article Title: Metabolic profiling and combined therapeutic strategies unveil the cytotoxic potential of selenium-chrysin (SeChry) in NSCLC cells

doi: 10.1042/BSR20240752

Figure Lengend Snippet: Analysis of the expression profiles of MCT1/SLC16A1 in normal lung tissue and primary carcinomas in ( A ) LUAD and ( B ) LUSC and MCT4/SLC16A3 in ( C ) LUAD and ( D ) LUSC extracted from the TCGA database. A two-tailed unpaired Student’s t -test with Welch’s correction was used. ( E ) Cells were kept in control conditions and immunofluorescence analysis showed that all cell lines express MCT1. Nuclei were stained with DAPI (blue). The white bars scale means 20 μm. The cytotoxic response to SeChry@PURE G4 -LA 24 combined with MCT1 or MCT4 inhibitors was evaluated. Cells were incubated overnight with the MCT1 inhibitor AZD3965 and/or the MCT4 inhibitor AZD0095 and the following day, they were treated with SeChry@PURE G4 -LA 24 at the determined LC50 concentration. Percentage of cell death analyzed by flow cytometry using Annexin V and PI in ( F ) A549, ( G ) H292, ( H ) PC-9 and ( I ) BEAS-2B cells. Data are represented as mean ± SD. * P <0.5, ** P <0.01, *** P <0.001, **** P <0.0001 (one-way ANOVA with Tukey’s multiple comparisons test was used). The effect of the combination of SeChry@PURE G4 -LA 24 with the EGFR inhibitor gefitinib was evaluated. Cells, ( J ) A549, ( K ) H292, ( L ) PC-9, ( M ) H522, ( N ) H596, ( O ) H1975, and ( P ) BEAS, were incubated overnight with gefitinib and the following day, they were treated with SeChry@PURE G4 -LA 24 at the determined LC50 concentration. Data are represented as mean ± SD. * P <0.5, ** P <0.01, *** P <0.001, **** P <0.0001 (one-way ANOVA with Tukey’s multiple comparisons test was used).

Article Snippet: Normal human bronchial epithelium cell line BEAS-2B (CRL-3588™, ATCC) was also used.

Techniques: Expressing, Two Tailed Test, Control, Immunofluorescence, Staining, Incubation, Concentration Assay, Flow Cytometry

Journal: PeerJ

Article Title: HtrA3: a promising prognostic biomarker and therapeutic target for head and neck squamous cell carcinoma

doi: 10.7717/peerj.16237

Figure Lengend Snippet: (A) Representative immunohistochemistry images of HtrA1-4 in HNSCC and normal tissues (HPA database); (B–C) protein expression of HtrA1 and HtrA3 in HNSCC and normal tissues with datasets from the CPTAC database; expression of HtrA1-4 in BEAS-2B, Cal-27 (D) and Fadu (E) cells obtained by western blot. Values associated with test proteins were normalized to standard β -actin for the relative expression measure; (F–G) column graphs of western blot analysis; (H) HtrA1-4 expression in BEAS-2B, FaDu, and Cal-27 cells by RT-PCR. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Human squamous carcinoma cell lines (Fadu and Cal-27) and human non-tumorigenic bronchial epithelium cells (BEAS-2B) (American Type Culture Collection, ATCC) were cultured in RPMI 1640 (Gibco BRL, Billings, MT, USA) and supplemented with 10% fetal bovine serum (FBS, Ex Cell) and antibiotics (100 U/Ml penicillin and 100 µg/Ml streptomycin) in a 5% CO2 incubator at 37 °C.

Techniques: Immunohistochemistry, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction