Journal:

Article Title: Airway epithelial cells synthesize the lipid mediator 5-oxo-ETE in response to oxidative stress

doi: 10.1016/j.freeradbiomed.2006.12.006

Figure Lengend Snippet: Epithelial cells used in this study

Article Snippet: Epithelial cells lines (A549, BEAS-2B, T84, and HEp-2) were obtained from the American Type Culture Collection ( ) and cultured in 10% FBS in DMEM/F12 medium containing L-glutamine (2 mM), sodium bicarbonate (2.5 g/L), penicillin (100U/ml), and streptomycin (100 μg/ml).

Techniques:

Journal:

Article Title: Airway epithelial cells synthesize the lipid mediator 5-oxo-ETE in response to oxidative stress

doi: 10.1016/j.freeradbiomed.2006.12.006

Figure Lengend Snippet: Epithelial cells were cultured to confluence in 6-well plates. Cells were washed twice with PBS+ and then preincubated for 6 min with 100 μM PMS. The cells were then incubated for an additional 20 min with 4 μM 5-HETE. Cells in adjoining wells were detached with trypsin and counted. Values are means ± SE (n = 4, except for T84 and HEp-2 cells (n=2))..

Article Snippet: Epithelial cells lines (A549, BEAS-2B, T84, and HEp-2) were obtained from the American Type Culture Collection ( ) and cultured in 10% FBS in DMEM/F12 medium containing L-glutamine (2 mM), sodium bicarbonate (2.5 g/L), penicillin (100U/ml), and streptomycin (100 μg/ml).

Techniques: Cell Culture, Incubation

Journal:

Article Title: Airway epithelial cells synthesize the lipid mediator 5-oxo-ETE in response to oxidative stress

doi: 10.1016/j.freeradbiomed.2006.12.006

Figure Lengend Snippet: Scheme showing the regulation of 5-oxo-ETE synthesis in airway epithelial cells. Metabolism of H2O2 and tBuOOH by glutathione peroxidase (GPerox) results in the formation of GSSG, which is reduced back to GSH by glutathione reductase (GRed), generating NADP+, which serves as a cofactor for 5-HEDH. This process can be blocked by NEM, which alkylates GSH, and mimicked by diamide (DA), which oxidizes GSH, and PMS, which oxidizes NADPH. In A549 cells the action of H2O2 is limited by metabolism by catalase, which can be blocked by azide.

Article Snippet: Epithelial cells lines (A549, BEAS-2B, T84, and HEp-2) were obtained from the American Type Culture Collection ( ) and cultured in 10% FBS in DMEM/F12 medium containing L-glutamine (2 mM), sodium bicarbonate (2.5 g/L), penicillin (100U/ml), and streptomycin (100 μg/ml).

Techniques:

Journal: PLoS ONE

Article Title: VBP15, a Glucocorticoid Analogue, Is Effective at Reducing Allergic Lung Inflammation in Mice

doi: 10.1371/journal.pone.0063871

Figure Lengend Snippet: A549 cells stably-transfected with a luciferase NFκB construct were exposed to increasing concentrations of VBP15 or prednisolone (3, 30, 300, 3000 nM) followed by TNFα stimulation before measuring luciferase activity. Bar graph represents mean (±SE) luciferase units. * p <.012 (due to the Bonferroni adjustment for multiple comparisons) compared to treatment with vehicle control. Data represents 4 biological replicates with assay performed in triplicate.

Article Snippet: Briefly, A549 epithelial cells (Panomics, Fremont, CA) stably transfected with a luciferase reporter construct regulated under NFκB response elements were grown in 75 mm 2 flasks according to the manufacturer's instructions and transferred to 96-well plates upon reaching confluency.

Techniques: Stable Transfection, Transfection, Luciferase, Construct, Activity Assay, Control