cell culture human alveolar epithelial type ii a549 cells  (Technology Development Co)

Journal: Acta Pharmacologica Sinica

Article Title: Paeoniflorin suppresses TGF-β mediated epithelial-mesenchymal transition in pulmonary fibrosis through a Smad-dependent pathway

doi: 10.1038/aps.2016.36

Figure Lengend Snippet: The effects of different concentrations of paeoniflorin (Pae) on TGF-β1-induced epithelial-mesenchymal transition in A549 cells. A549 cells were seeded into plates and incubated with serum-free RPMI 1640 for 2 h. Then, the cells were incubated with TGF-β1 (2 ng/mL) or TGF-β1 (2 ng/mL) plus Pae (1, 3, 10 and 30 μmol/L) for the indicated intervals. (A) The expression levels of E-cadherin, Vimentin and α-SMA were detected using Western blot analysis. (B) The amounts of type I and type III collagen in the cell supernatants were detected using ELISA and Real-Time Quantitative PCR assays. Data are expressed as the means±SD. n=3. ##P<0.01 vs normal. *P<0.05, **P<0.01 vs TGF-β1.

Article Snippet: Cell culture Human type II alveolar epithelial cell lines (A549), obtained from the American Type Culture Collection (ATCC), were cultured in RPMI-1640 containing 10% FBS, 100 units/mL penicillin, 100 μg/mL streptomycin and maintained at 37 °C in a 5% CO 2 humidified atmosphere.

Techniques: Incubation, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Acta Pharmacologica Sinica

Article Title: Paeoniflorin suppresses TGF-β mediated epithelial-mesenchymal transition in pulmonary fibrosis through a Smad-dependent pathway

doi: 10.1038/aps.2016.36

Figure Lengend Snippet: Time course of the effect of paeoniflorin (Pae) on TGF-β1-induced epithelial-mesenchymal transition in A549 cells. A549 cells were seeded into plates and incubated with serum-free RPMI-1640 for 2 h. Then, the cells were incubated with TGF-β1 (2 ng/mL) or TGF-β1 (2 ng/mL) plus Pae (1, 3, 10 and 30 μmol/L) for the indicated intervals. (A) Cell morphology was recorded using an inverted microscope (magnification 200×). (B) The expression levels of E-cadherin, Vimentin and α-SMA were detected using Western blot analysis, as described in the Methods section. GAPDH was used as the internal control. (C) The protein levels of E-cadherin, Vimentin and α-SMA were determined using Real-Time Quantitative PCR, as described in the Methods section. (D) The cell supernatants were collected, and the amount of type I collagen was detected using ELISA, as described in the Methods section. The data are expressed as the mean±SD. n=3. ##P<0.01 vs normal. *P<0.05, **P<0.01 vs TGF-β1.

Article Snippet: Cell culture Human type II alveolar epithelial cell lines (A549), obtained from the American Type Culture Collection (ATCC), were cultured in RPMI-1640 containing 10% FBS, 100 units/mL penicillin, 100 μg/mL streptomycin and maintained at 37 °C in a 5% CO 2 humidified atmosphere.

Techniques: Incubation, Inverted Microscopy, Expressing, Western Blot, Control, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Cell

Article Title: High-content siRNA screen reveals global ENaC regulators and potential cystic fibrosis therapy targets.

doi: 10.1016/j.cell.2013.08.045

Figure Lengend Snippet: Figure 1. ENaC Functional Screening Assay (A) Images of A549 cells subjected to the FMP/ Amil assays before (left) and after Amil (right). Cells grown for 48 hr under dexamethasone (+Dexa, top) demonstrated enhanced FMP/Amil quench- ing (higher ENaC activity) than cells grown in the absence of Dexa (bottom). (B) Quantification of the Amil-sensitive FMP fluo- rescence of images in A shows greater ENaC activity in cells grown under Dexa. (C) Images of A549 cells grown for 48 hr under Scrbld, aENaC, or ßCOP siRNAs. (D) Summary of the median ratio Amil-sensitive FMP fluorescence ratios for all spots in the same slide (‘‘slide’’), scrambled (‘‘Scrbld’’), aENaC and ßCOP siRNA spots. Average of Amil-sensitive ra- tios (n = 5) is shown as a red line ± SD.

Article Snippet: Cell Culture Human alveolar type II epithelial A549 cells (ATCC, Cat no. CCL-185) were grown with or without dexamethasone (Dexa) as before, as well as WT- or F508del-CFTR transduced A549 cells (Almaça et al., 2011).

Techniques: Functional Assay, Screening Assay, Activity Assay

Journal: Journal of Interferon & Cytokine Research

Article Title: Inhibition by New Glucocorticoid Antedrugs [16α, 17α-d] Isoxazoline and [16α, 17α-d]-3′-Hydroxy-Iminoformyl Isoxazoline Derivatives of Chemotaxis and CCL26, CCL11, IL-8, and RANTES Secretion

doi: 10.1089/jir.2012.0129

Figure Lengend Snippet: Table 1.

Article Snippet: The structures of these compounds are shown in . fig ft0 fig mode=article f1 FIG. 1. caption a4 Structures of glucocorticoid antedrugs (GCAs). table ft1 table-wrap mode=article t1 Table 1. caption a4 Name and Nomenclature for Glucocorticoid Antedrugs Cell culture Human A549 alveolar type II epithelial-like cells (HAE cells) [American Type Culture Collection (ATCC) CCL-185] were grown in RPMI-1640/F-12K (50:50 vol/vol) supplemented with 10% fetal calf serum, penicillin (100 U/mL), and streptomycin (100 μg/mL) in a humidified atmosphere of 5% carbon dioxide at 37°C.

Techniques: