Figure 1.

Effects of citrinin in BIO-treated ESCs and ES-β-catenin clones. (A, B): ES cells (1 × 10⁴ cells) without and with BIO treatment for the indicated times and (C, D) ES β-catenin clones. To evaluate the effects of β-catenin accumulation in the control of apoptosis, ES BIO-treated cells and ES β-catenin clones were then treated with citrinin (60 μM) for 24 hours. (A, C): Apoptosis was evaluated using Cell Death Detection Elisa kits (mean ± SEM; n = 3). (B, D): Cell viability was determined by quantitation of ATP with the Cell Titer-Glo Luminescent Cell Viability Assay (mean ± SEM; n = 3). Fold changes were calculated with respect to No BIO samples. (E): ES β-catenin clones were analyzed for proliferation potential using the FACS-CFSE system (mean ± SEM; n = 3). (F): Quantitative RT-PCR analysis of p16 and p19 transcript levels in the ES-β-catenin clones (mean ± SEM; n = 3). Abbreviations: BIO, 6-bromoindirubin-3′-oxime; CFSE, 5-(and 6-) carboxyfluorescein diacetate succinimidyl ester; ES, embryonic stem; FACS, fluorescence activated cell sorting; RT-PCR, real time PCR; wt, wild type.