Journal: International Journal of Molecular Sciences

Article Title: Co-Stimulation with TWEAK and TGF-β1 Induces Steroid-Insensitive TSLP and CCL5 Production in BEAS-2B Human Bronchial Epithelial Cells

doi: 10.3390/ijms252111625

Figure Lengend Snippet: Molecular mechanisms resulting in steroid unresponsiveness, induced by co-stimulation of TWEAK and TGF-β1. The mRNA levels of MKP-1 ( A ), GILZ ( C ), GRβ ( E ), and HDAC2 ( F ) mRNA after 48 h treatment, analyzed by qRT–PCR. MAP kinase phosphatase 1 (MKP-1) and glucocorticoid-induced leucine zipper (GILZ) levels analyzed by immunoblotting ( B , D , upper panel). Densitometric analysis of protein bands ( B , D , lower). Data represent mean ± SD of two independent experiments. * p < 0.05, compared to untreated cultures as controls; † p < 0.05 compared with cultures in the absence of DEX.

Article Snippet: Anti-MAPK phosphatase-1 (MKP-1) monoclonal antibody (sc-271684) and anti-glucocorticoid-induced leucine zipper (GILZ) mAb (G-5) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Co-Stimulation with TWEAK and TGF-β1 Induces Steroid-Insensitive TSLP and CCL5 Production in BEAS-2B Human Bronchial Epithelial Cells

doi: 10.3390/ijms252111625

Figure Lengend Snippet: Co-stimulation of TWEAK and TGF-β1 induces steroid-unresponsive production of cytokines through the mitogen-activated protein kinase and nuclear factor kappa beta signaling pathways. Confluent monolayers of BEAS-2B cells were cultured for 48 h in the absence (DMSO as vehicle) or presence of AZD6244 (5 μM), SB202190 (5 μM), SP600125 (5 μM), or BAY11-7082 (2.5 μM) and treated with TGF-β1 (10 ng/mL), TWEAK (100 ng/mL), or TGF-β1 in combination with TWEAK. The mRNA levels of TSLP ( A ), CCL5 ( B ), MKP-1 ( C ), and GILZ ( D ) were analyzed by qRT–PCR. Data represent mean ± SD of two independent experiments. * p < 0.05, compared with the vehicle; † p < 0.05 compared with cultures in the absence of DEX.

Article Snippet: Anti-MAPK phosphatase-1 (MKP-1) monoclonal antibody (sc-271684) and anti-glucocorticoid-induced leucine zipper (GILZ) mAb (G-5) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Protein-Protein interactions, Cell Culture, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Co-Stimulation with TWEAK and TGF-β1 Induces Steroid-Insensitive TSLP and CCL5 Production in BEAS-2B Human Bronchial Epithelial Cells

doi: 10.3390/ijms252111625

Figure Lengend Snippet: MKP-1 is involved in steroid unresponsiveness induced by co-stimulation with TWEAK and TGF-β1 in BEAS-2B cells. The mRNA levels of MKP-1 ( A ), TSLP ( C ), and CCL5 ( E ), analyzed by qRT–PCR. Data represent mean ± SD of two independent experiments. MKP-1 and thymic stromal lymphopoietin (TSLP) expression after 48 h of treatment, assessed by immunoblotting ( B , D , upper panel). Densitometric analysis of protein bands ( B , D , lower). The level of CCL5 in cell culture supernatants after 48 h of treatment, analyzed using ELISA ( F ). Data represent mean ± SD of two independent experiments. * p < 0.05, compared to untreated cultures as controls; † p < 0.05 compared with cultures in the absence of DEX; ‡ p < 0.05 compared with control siRNA.

Article Snippet: Anti-MAPK phosphatase-1 (MKP-1) monoclonal antibody (sc-271684) and anti-glucocorticoid-induced leucine zipper (GILZ) mAb (G-5) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Control