Fig. 6. Ectopic expression of p65 ablates TGF-ß1-mediated growth arrest of Hs578T cells. Hs578T cells were plated in triplicate at 70% confluence in 96-well dishes. After removal of the media, cells were incubated according to the manufacturers directions for 24 h in a 4-µl solution of DNA in FUGENE [either 130 ng pMT2T parental + 20 ng GFP/well, or 130 ng of human p65 pMT2T + 20 ng GFP/well (+p65)]. After 24 h, the cells were treated with carrier BSA (B) or with 5 ng/ml TGF-ß1 (T), and the effects of TGF-ß1 on growth were measured by MTS assay. The average of two experiments are shown; values are given as percent cell proliferation relative to BSA carrier-treated control cells that had been transfected with the parental pMT2T vector DNA. Transfection efficiency was estimated to be approximately 70% based on GFP staining.