Figure 1

Transient transfection of GFP-BAX fusion protein into HCT116 ^{BAX -/-} cells rescues cell death after Indomethacin treatment. (A) HCT116^BAX-/-cells are resistant to indomethacin. Line graph showing percentage of apoptotic cells in the medium after exposure of 500 μM indomethacin for wild type (WT) and BAX-deficient HCT116 cells over time. (B) Rescue of the cell death phenotype by transfection of an exogenous GFP-Bax gene. HCT116^{BAX -/-} cells were transfected with either no plasmid, pEGFP-C3 (GFP), or pEGFP-C3-BAX (GFP-BAX), which expresses a fusion protein of GFP in phase with the murine Bax coding region under the control of the CMV immediate early promoter. Twenty-four hours after transfection, cells were treated with 500 μM indomethacin (+Indo) or vehicle (-Indo). Transfection efficiency was ~60% in each condition, based on the proportion of cells positive for GFP expression. Forty-eight hours after indomethacin treatment, cell death was assessed. Transfection of GFP-BAX into cells that were later treated with indomethacin caused a significant increase in death over GFP transfection and indomethacin-treated cells (t-test, *P = 0.0001). Values shown are mean ± SD in both graphs of data collected from 3 independent experiments.