Cells were serum-starved for 24 h and then treated with 5 ng/mL porcine transforming growth factor beta 1 (TGF-beta, R&D Systems, Minneapolis, MN) for indicated times.
Growth protocol
A549 cells were maintained in RPMI-1640 medium with glutamine, supplemented with 10% FBS, penicillin, and streptomycin and tested for mycoplasma contamination. All tissue culture media and media supplements were purchased from Life Technologies (Gaithersburg, MD).
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated from the harvested cells using Trizol (Invitrogen) and cleanup performed with an RNeasy kit (Qiagen)
Label
biotin
Label protocol
Biotinylated cRNA was prepare from 3-5 ug total RNA using standard Affymetrix protocol