FOXO3A induces growth arrest. (A) FOXO3A inhibits growth of cHL cell lines. Cells transfected with a vector containing FOXO3(A3)ER (F3ER) or EV were seeded in 6-well plates at a density of 0.05 × 106 cells per mL. Twenty-four hours later, cells were treated with 4-OHT or vehicle. Live cells were counted with a hemacytometer at indicated time points after seeding. Data represent mean ± SD of 3 independent experiments. (B) F3ER- and EV-expressing cells were treated or not treated with 200 nM 4-OHT. Cell death was examined at indicated time points by staining with Annexin V/PI. Specific cell death was calculated as described in “Material and methods” and is shown as mean ± SD of 3 independent experiments. (C) FOXO3A activation decreases the proportion of cells in the S phase. A total of 1 × 106 cells were seeded in 10 mL of complete medium and treated with 4-OHT or with vehicle. After 24 hours, cells were harvested and cell cycle was analyzed by PI staining. Bars represent mean ± SD of 3 independent experiments. Significance was calculated using a 2-sided t test. *P < .05; **P < .01; ***P < .001; ****P < .0001.
