bronchial epithelial cell beas2b tissue culture  (ATCC)

Journal: FASEB BioAdvances

Article Title: Endoplasmic Reticulum Stress and Unfolded Protein Response Sensor ERN1 Regulates Organic Dust Induction of Lung Inflammation

doi: 10.1096/fba.2025-00069

Figure Lengend Snippet: Time‐course effects of dust extract on the expression of ER stress‐UPR sensors in Beas2B bronchial epithelial cells. Cells were treated with 1% dust extract for 3, 9, and 24 h and protein and mRNA levels of ER stress‐UPR sensors were determined. (A) Representative western blots are shown. (B–E) HSPA5, ERN1, EIF2AK3 and ATF6 protein levels were determined by western blotting and normalized to Actin levels. (F–I) HSPA5, ERN1, EIF2AK3, and ATF6 mRNA levels were determined by real‐time qRT‐PCR and normalized to Actin mRNA levels. Data shown are mean ± SE ( n = 3–4). Statistical significance was analyzed by one‐way ANOVA followed by Dunnett's post hoc test. * p < 0.05, ** p < 0.01. C, control; DE, dust extract.

Article Snippet: Beas2B bronchial epithelial cells (ATCC CRL‐9609, Manassas, VA, USA) were grown on plastic culture dishes coated with fibronectin, bovine type 1 collagen and bovine serum albumin and maintained in LHC 9 (12680013, Thermo Fisher Scientific, Waltham, MA, USA) or BronchiaLife (LS‐1047, Lifeline Cell Technology, Oceanside, CA, USA) medium containing penicillin (100 U/mL), streptomycin (100 μg/mL), gentamicin (30 μg/mL) and amphotericin B (0.25 μg/mL).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Control

Journal: FASEB BioAdvances

Article Title: Endoplasmic Reticulum Stress and Unfolded Protein Response Sensor ERN1 Regulates Organic Dust Induction of Lung Inflammation

doi: 10.1096/fba.2025-00069

Figure Lengend Snippet: Effects of dust extract on ERN1 phosphorylation (Ser724) and XBP‐1 splicing in Beas2B bronchial epithelial cells and mouse lungs. (A, B) Cells were treated with 1% dust extract (DE) for 5, 10, 30, 60, and 120 min, and the levels of phosphorylated and total ERN1 were detected by western blotting. Levels of phospho ERN1 were normalized to total ERN1. Data shown are mean ± SE ( n = 3). A representative western blot is shown (A). (C, D) Cells were treated with 1% dust extract for 3, 9, and 24 h. Unspliced (XBP1u) and spliced (XBP1s) protein levels were determined by western blotting and normalized to Actin levels. A representative western blot is shown (C). Data shown are mean ± SE ( n = 4). Statistical significance for (B, D) was analyzed by one‐way ANOVA followed by Dunnett's post hoc test. C, control; DE, dust extract. (E–H) Lung sections from mice ( n = 4) repeatedly treated with dust extract, as indicated in Figure legend, were immunostained with phosphorylated ERN1 (pIRE) and non‐immune rabbit IgG antibodies and developed with DAB substrate. The immunohistochemical images were blinded. Representative images are shown at 10 and 40× magnifications (Scale bar = 500 μm).

Article Snippet: Beas2B bronchial epithelial cells (ATCC CRL‐9609, Manassas, VA, USA) were grown on plastic culture dishes coated with fibronectin, bovine type 1 collagen and bovine serum albumin and maintained in LHC 9 (12680013, Thermo Fisher Scientific, Waltham, MA, USA) or BronchiaLife (LS‐1047, Lifeline Cell Technology, Oceanside, CA, USA) medium containing penicillin (100 U/mL), streptomycin (100 μg/mL), gentamicin (30 μg/mL) and amphotericin B (0.25 μg/mL).

Techniques: Phospho-proteomics, Western Blot, Control, Immunohistochemical staining

Journal: FASEB BioAdvances

Article Title: Endoplasmic Reticulum Stress and Unfolded Protein Response Sensor ERN1 Regulates Organic Dust Induction of Lung Inflammation

doi: 10.1096/fba.2025-00069

Figure Lengend Snippet: Effects of TLR chemical inhibitors and MyD88 knockdown on dust extract induction of ERN1 protein levels in Beas2B bronchial epithelial cells. (A–D) Cells were treated with medium alone, 10 μM CU CPT 22, or 10 μM resatorvid for 1 h prior to incubation with 1% dust extract for 24 h. (E–G) Cells were transfected with 10 nM non‐targeting control siRNA or MyD88 siRNA and treated with 1% dust extract for 24 h. ERN1 and MyD88 protein levels were determined by western blotting and normalized to Actin levels. Representative western blots are shown (A, C, E). Data shown are mean ± SE ( n = 4 for chemical inhibitors and n = 3 for MyD88 siRNA transfection). Statistical significance was analyzed by two‐way ANOVA followed by Sidak's post hoc test for analysis between specific groups (F) and Tukey's post hoc test for analysis between multiple groups (B, D, G). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. C, control; DE, dust extract.

Article Snippet: Beas2B bronchial epithelial cells (ATCC CRL‐9609, Manassas, VA, USA) were grown on plastic culture dishes coated with fibronectin, bovine type 1 collagen and bovine serum albumin and maintained in LHC 9 (12680013, Thermo Fisher Scientific, Waltham, MA, USA) or BronchiaLife (LS‐1047, Lifeline Cell Technology, Oceanside, CA, USA) medium containing penicillin (100 U/mL), streptomycin (100 μg/mL), gentamicin (30 μg/mL) and amphotericin B (0.25 μg/mL).

Techniques: Knockdown, Incubation, Transfection, Control, Western Blot

Journal: FASEB BioAdvances

Article Title: Endoplasmic Reticulum Stress and Unfolded Protein Response Sensor ERN1 Regulates Organic Dust Induction of Lung Inflammation

doi: 10.1096/fba.2025-00069

Figure Lengend Snippet: Effect of NOX chemical inhibitor VAS2870 on dust extract induction of ERN1 protein levels in Beas2B bronchial epithelial cells. Cells were treated with medium alone or 5 μM VAS2870 for 1 h prior to incubation with 1% dust extract for 24 h. ERN1 protein levels were determined by western blotting and both ERN1 minor upper band and major lower band were quantified and normalized to Actin levels. A representative western blot is shown with noncontiguous lanes demarcated by broken lines (A). Data shown are mean ± SE ( n = 4) and statistical significance was analyzed by two‐way ANOVA followed by Tukey's post hoc test (B). ** p < 0.01. C, control; DE, dust extract.

Article Snippet: Beas2B bronchial epithelial cells (ATCC CRL‐9609, Manassas, VA, USA) were grown on plastic culture dishes coated with fibronectin, bovine type 1 collagen and bovine serum albumin and maintained in LHC 9 (12680013, Thermo Fisher Scientific, Waltham, MA, USA) or BronchiaLife (LS‐1047, Lifeline Cell Technology, Oceanside, CA, USA) medium containing penicillin (100 U/mL), streptomycin (100 μg/mL), gentamicin (30 μg/mL) and amphotericin B (0.25 μg/mL).

Techniques: Incubation, Western Blot, Control

Journal: FASEB BioAdvances

Article Title: Endoplasmic Reticulum Stress and Unfolded Protein Response Sensor ERN1 Regulates Organic Dust Induction of Lung Inflammation

doi: 10.1096/fba.2025-00069

Figure Lengend Snippet: Effects of NFκB and Stat3 chemical inhibitors and Stat3 knockdown on dust extract induction of ERN1 protein levels in Beas2B bronchial epithelial cells. (A–D) Cells were first treated for 1 h with medium alone, 5 μM BAY 11–7082 (BAY), or 5 μM Stattic prior to incubation with 1% dust extract for 24 h. (E–G). Cells were transfected with 33 nM non‐targeting control siRNA or Stat3 siRNA and treated with 1% dust extract for 24 h. ERN1 and Stat3 protein levels were determined by western blotting and normalized to Actin levels. In (A, C), both ERN1 minor upper band and major lower band were quantified and normalized to Actin. Representative western blots are shown with noncontiguous lanes demarcated by broken lines (A, C). Data shown are mean ± SE (B: n = 6; D: n = 4; F: n = 4–5; G: n = 5). Statistical significance was analyzed by two‐way ANOVA followed by Sidak's post hoc test for analysis between specific groups (F) and Tukey's post hoc test for analysis between multiple groups (B, D, G). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. C, control; DE, dust extract.

Article Snippet: Beas2B bronchial epithelial cells (ATCC CRL‐9609, Manassas, VA, USA) were grown on plastic culture dishes coated with fibronectin, bovine type 1 collagen and bovine serum albumin and maintained in LHC 9 (12680013, Thermo Fisher Scientific, Waltham, MA, USA) or BronchiaLife (LS‐1047, Lifeline Cell Technology, Oceanside, CA, USA) medium containing penicillin (100 U/mL), streptomycin (100 μg/mL), gentamicin (30 μg/mL) and amphotericin B (0.25 μg/mL).

Techniques: Knockdown, Incubation, Transfection, Control, Western Blot

Journal: FASEB BioAdvances

Article Title: Endoplasmic Reticulum Stress and Unfolded Protein Response Sensor ERN1 Regulates Organic Dust Induction of Lung Inflammation

doi: 10.1096/fba.2025-00069

Figure Lengend Snippet: Effects of ERN1 chemical inhibitors and ERN1 knockdown on dust extract induction of inflammatory mediator protein levels in Beas2B bronchial epithelial cells. (A–E). Cells were first treated for 1 h with medium alone, 1 μM KIRA6, or 1 μM APY29 prior to incubation with 1% dust extract for 3 h. (F–K). Cells were transfected with 10 nM non‐targeting control siRNA or ERN1 siRNA and treated with 1% dust extract for 24 h. Cellular pro IL1β, ICAM1, and ERN1 protein levels and secreted IL6 and CXCL8 levels were determined by western blotting and ELISA, respectively. Pro IL1β, ICAM1, and ERN1 protein levels were normalized to Actin levels. Representative western blots are shown (A, F). Data shown are mean ± SE ( n = 4–8 for chemical inhibitor experiments and n = 4–5 for siRNA transfection experiments). Statistical significance was analyzed by two‐way ANOVA followed by Sidak's post hoc test for analysis between specific groups (G) and Tukey's post hoc test for analysis between multiple groups (B–E, H–K). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. C, control; DE, dust extract.

Article Snippet: Beas2B bronchial epithelial cells (ATCC CRL‐9609, Manassas, VA, USA) were grown on plastic culture dishes coated with fibronectin, bovine type 1 collagen and bovine serum albumin and maintained in LHC 9 (12680013, Thermo Fisher Scientific, Waltham, MA, USA) or BronchiaLife (LS‐1047, Lifeline Cell Technology, Oceanside, CA, USA) medium containing penicillin (100 U/mL), streptomycin (100 μg/mL), gentamicin (30 μg/mL) and amphotericin B (0.25 μg/mL).

Techniques: Knockdown, Incubation, Transfection, Control, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: FASEB BioAdvances

Article Title: Endoplasmic Reticulum Stress and Unfolded Protein Response Sensor ERN1 Regulates Organic Dust Induction of Lung Inflammation

doi: 10.1096/fba.2025-00069

Figure Lengend Snippet: Effects of ERN1 chemical inhibitor KIRA6 on dust extract induced NFκB‐p65, Stat3, Jun and MAPK 8/9 phosphorylation in Beas2B bronchial epithelial cells. Cells were first treated for 1 h with medium alone or 1 μM KIRA6. After treatment, cells were incubated with medium alone or 1% dust extract for 10 min to analyze NFκB‐p65 phosphorylation, 1 h to analyze Stat3 and Jun phosphorylation, and 20 min to analyze MAPK 8/9 phosphorylation. Phosphorylated and total levels of NFκB‐p65, Stat3, Jun and MAPK 8/9 were determined by western blotting and the levels of phosphorylated proteins were normalized to total protein levels. Representative western blots are shown (A, C, E and G). Data shown are mean ± SE (B and D: N = 5; F and H: N = 6). Statistical significance was analyzed by two‐way ANOVA followed by Tukey's post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001. C, control; DE, dust extract.

Article Snippet: Beas2B bronchial epithelial cells (ATCC CRL‐9609, Manassas, VA, USA) were grown on plastic culture dishes coated with fibronectin, bovine type 1 collagen and bovine serum albumin and maintained in LHC 9 (12680013, Thermo Fisher Scientific, Waltham, MA, USA) or BronchiaLife (LS‐1047, Lifeline Cell Technology, Oceanside, CA, USA) medium containing penicillin (100 U/mL), streptomycin (100 μg/mL), gentamicin (30 μg/mL) and amphotericin B (0.25 μg/mL).

Techniques: Phospho-proteomics, Incubation, Western Blot, Control