normal human bronchial epithelial cell line beas 2b  (ATCC)

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: Cell Research

Article Title: Mucus production stimulated by IFN-AhR signaling triggers hypoxia of COVID-19

doi: 10.1038/s41422-020-00435-z

Figure Lengend Snippet: a hACE2-transgenic mice were infected with SARS-CoV-2 for the indicated time period. The expression of IFN-β was measured by real-time PCR. b Immunohistochemical staining of IFN-β from the lung sections of SARS-CoV-2-infected hACE2-transgenic mice for 5 days. Scale bars, 50 μm. c Representative image of immunohistochemical staining of isotype control-IgG or IFN-β in BAL from three COVID-19 patients. Scale bars, 50 μm. d BEAS-2B cells were treated with different doses of IFN-β (1 or 10 ng/mL) for 24 h. The expression of secreted mucins ( MUCs 2 , 5A , 5B , 6 , 9 and 19 ) and membrane-mucins ( MUCs 1 , 7 , 12 , 15 , 16 , 17 , 18 and 20 ) was determined by real-time PCR. e Western blot analysis of the expression of mucins 5A, 5B, 7 and 16 from BEAS-2B cells treated with PBS, IFN-β (1 ng/mL) or IFN-γ (10 ng/mL) for 48 h. f The same as ( d ), except that primary alveolar epithelial cells (PAECs) were treated with IFN-β (1 ng/mL) or IFN-γ (10 ng/mL). g Institute of Cancer Research (ICR) mice were treated with IFN-β (1 μg/mouse) or IFN-γ (10 μg/mouse) through the trachea once every day for 4 days. The expression of mucins 1, 2, 5A, 5B, 6 and 13 from the lung tissues was determined by western blot. h The same as ( g ), except the mucus was shown from the tracheal tubes. White arrow indicated the mucus. The data represent means ± SD. Representative images are from three mice ( b , e , g and h ), four mice ( a ) or three independent experiments ( d and f ). * P < 0.05, ** P < 0.01, *** P < 0.001, by one-way ANOVA ( a , d and f ) or two-tailed Student’s t -test ( b ).

Article Snippet: Normal human bronchial epithelium cell line BEAS-2B cells were purchased from the China Center for Type Culture Collection (Shanghai, China) and cultured in RPMI1640 medium (Gibco, USA) or DMEM medium (Gibco, USA) with 10% FBS.

Techniques: Transgenic Assay, Infection, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Western Blot, Two Tailed Test

Journal: Cell Research

Article Title: Mucus production stimulated by IFN-AhR signaling triggers hypoxia of COVID-19

doi: 10.1038/s41422-020-00435-z

Figure Lengend Snippet: a , b hACE2-transgenic mice were infected with SARS-CoV-2. The expression of IFN-γ was detected at day 7 after SARS-CoV-2 infection by real-time PCR ( a ) or immunohistochemical staining with anti-IFN-γ antibody ( b ). Scale bars, 50 μm. c BEAS-2B cells were treated with different doses of IFN-γ (1 or 10 ng/mL) for 24 h. The expression of secreted mucins ( MUCs 2 , 5A , 5B , 6 , 9 and 19 ) and membrane-mucins ( MUCs 1 , 7 , 12 , 15 , 16 , 17 , 18 and 20 ) was determined by real-time PCR. d The same as ( b ), except that the expression of mucins 1, 2 and 5A was determined. Scale bars, 50 μm. The data represent means ± SD. Representative images are from three mice ( b and d ), four mice ( a ) or three biological independent samples ( c ). ** P < 0.01, *** P < 0.001, by two-tailed Student’s t -test ( a , b and d ) or one-way ANOVA ( c ).

Article Snippet: Normal human bronchial epithelium cell line BEAS-2B cells were purchased from the China Center for Type Culture Collection (Shanghai, China) and cultured in RPMI1640 medium (Gibco, USA) or DMEM medium (Gibco, USA) with 10% FBS.

Techniques: Transgenic Assay, Infection, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Two Tailed Test

Journal: Cell Research

Article Title: Mucus production stimulated by IFN-AhR signaling triggers hypoxia of COVID-19

doi: 10.1038/s41422-020-00435-z

Figure Lengend Snippet: a PAECs or BEAS-2B cells were treated with IFN-β (1 ng/mL) or IFN-γ (10 ng/mL) for 48 h. The expression of IDO1 was determined by western blotting. b BEAS-2B cells were treated with IFN-β (1 ng/mL) or IFN-γ (10 ng/mL) for 48 h. The levels of Kyn from cell lysates or the supernatants were determined by ELISA. c PAECs were isolated from AhR –/– mice were treated with IFN-β (1 ng/mL) or IFN-γ (10 ng/mL) for 24 h. The expression of Mucs 1 , 2 , 5A , 6 , 13 , 15 and 16 was determined by real-time PCR. d BEAS-2B cells were treated with Kyn (400 μM) for 24 h. The expression of mucins ( MUCs 2 , 5A , 6 , 7 , 13 , 15 , 16 , 17 , 18 and 20 ) was determined by real-time PCR. e Western blot analysis of the expression of mucins 1, 5A, 5B, 13 and 16 from BEAS-2B cells or PAECs treated with Kyn (400 μM) for 48 h. The data represent means ± SD. In b – d , n = 3 biological independent samples. Representative images are from three independent experiments ( a and e ). * P < 0.05, ** P < 0.01, *** P < 0.001, by two-tailed Student’s t -test ( d ) or one-way ANOVA ( b and c ).

Article Snippet: Normal human bronchial epithelium cell line BEAS-2B cells were purchased from the China Center for Type Culture Collection (Shanghai, China) and cultured in RPMI1640 medium (Gibco, USA) or DMEM medium (Gibco, USA) with 10% FBS.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Isolation, Real-time Polymerase Chain Reaction, Two Tailed Test

Journal: Cell Research

Article Title: Mucus production stimulated by IFN-AhR signaling triggers hypoxia of COVID-19

doi: 10.1038/s41422-020-00435-z

Figure Lengend Snippet: a , b BEAS-2B cells ( a ) or PAECs ( b ) were treated with IFN-β (1 ng/mL), IFN-γ (10 ng/mL) or Kyn (400 μM) for 48 h. Cells were immunostained with an anti-AhR antibody and observed under a confocal microscope. Scale bar, 10 μm. c BEAS-2B cells were treated with IFN-β (1 ng/mL), IFN-γ (10 ng/mL) or Kyn (400 μM) for 48 h. ChIP-qPCR analysis was performed with anti-AhR antibody and MUCs 2, 5A, 5B, 1, 13 and 16 promoter-specific primers. Data are present as relative fold enrichment to the PBS group. d hACE2-transgenic mice were infected with SARS-CoV-2 and the expression and localization of AhR (red) from the lung tissues was observed under confocal microscope. Scale bar, 10 μm. The data represent means ± SD. In c , n = 3 biological independent samples. Representative images are from three independent experiments ( a and b ) or three mice ( d ). * P < 0.05, ** P < 0.01, *** P < 0.001, by two-tailed Student’s t -test ( d ) or one-way ANOVA ( c ).

Article Snippet: Normal human bronchial epithelium cell line BEAS-2B cells were purchased from the China Center for Type Culture Collection (Shanghai, China) and cultured in RPMI1640 medium (Gibco, USA) or DMEM medium (Gibco, USA) with 10% FBS.

Techniques: Microscopy, Transgenic Assay, Infection, Expressing, Two Tailed Test