NCI-H292 cells (5 × 105 cells/well) were seeded in 6-well plates. The cells were incubated for 12 h in GM, and the medium was subsequently changed to serum-reduced GM medium (0.1% FBS). After 16 h, the cells were treated with the respective concentration of TGFβ1 for 30 min. Proteins were prepared and loaded as described elsewhere (Lee et al., 2014 (link)). At least 30 µg of protein from the whole cell lysate was used per sample for western blot analysis. The band intensity was visualized using a LAS-4000 luminescent image analyzer (Fujifilm, Japan) and quantified by densitometry (Fuji Multi Gauge software ver. 3.0).
Protocol detail
TGFβ1-Induced Protein Expression in NCI-H292 Cells
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Corresponding Organization :
Other organizations : Korea Research Institute of Bioscience and Biotechnology, Chungnam National University, Korea University of Science and Technology, Chungnam National University Hospital
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Variable analysis
independent variables
- TGFβ1 concentration
- Protein expression (band intensity)
- NCI-H292 cell line
- Seeding density (5 × 10^5 cells/well)
- Incubation time (12 h in GM, 16 h in serum-reduced GM)
- Protein extraction and loading procedure
- Western blot analysis (LAS-4000 luminescent image analyzer, Fuji Multi Gauge software ver. 3.0)
- Positive control: Not specified
- Negative control: Not specified
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