Sw 1573 cells

A549, A375, HEK-293T, HeLa, MDA-MB-231, RKO, and HepG2 cells (American Type Culture Collection, Rockville, MD) and 293GP2 packaging cells (ATTC) were cultured in DMEM (Gibco) supplemented with 10% FBS (Gibco). MCF10a cells (ATCC) were cultured in DMEM supplemented with horse serum (5%), EGF (20ng/ml), Hydrocortisone (0.5 mg/ml), Cholera Toxin (100 ng/ml) and Insulin (10 μg/ml). MIA PaCa-2 cells (ATCC) were cultured in DMEM with 10% fetal bovine serum and 2.5% horse serum (Gibco). SW-1573 cells (ATCC) were grown in RPMI-1640 (Gibco) with 10% fetal bovine serum. NIH3T3 and 3T3^RAS cells were grown in DMEM with 10% calf serum (Colorado Serum Company). All media were supplemented with Normocin (InvivoGen) to prevent Mycoplasma contamination. Cell lines were not authenticated. Cells were generally passaged fewer than 5 times, and freshly thawed cells were maintained in culture for no more than two weeks before conducting experiments. For serum starvation, cells were plated and cultured for 24 hr, the media was replaced by DMEM containing 0.1% serum for 24 hr, after which the cells were stimulated with 10% serum and harvested at the indicated time points.