Fig. 6

FOXO3-GFP translocation requires the p110α subunit of PI3Kinase and transactivation of the EGFR. a FOXO3-GFP-transfected MCF7 cells were serum starved for 24 h prior to the following treatments. Cells were treated with either 50 nM estrogen, 100 nM G-1 or 50 ng/ml EGF for 15 min; where indicated cells were pretreated with 10 μM LY294002 (a broad PI3Kinase inhibitor), 250 nM AG1478 (EGFR inhibitor) or vehicle (DMSO) for 30 min. b Cells were treated with estrogen, G-1 or EGF as in (a) following pretreatment with 100 nM PIK-75 (a p110α-selective inhibitor), 100 nM TGX-221 (a p110β-selective inhibitor) or vehicle (DMSO) for 30 min as indicated. For (a) and (b), results are reported as mean +/− s.e.m. from at least 3 experiments. *, p < 0.05; **, p < 0.01 vs. DMSO control. #, p < 0.05 vs. the (ligand-matched) DMSO treatment. c MCF7 cells were treated with 100 nM PIK-75, 100 nM TGX-221 or a combination of both inhibitors, in the absence of any ligand, as in (b). Results are reported as mean +/− s.e.m. from 3 experiments. *, p < 0.05 vs. DMSO control. #, p < 0.05 vs. treatment with TGX alone