Time-course experiments of cultured thymocytes. (A) Western blot analysis of GILZ (top), Bcl-xL (middle), and β-tubulin (bottom) proteins expressed in WT or TG thymocytes freshly isolated (0 h) or after 30 minutes, 1 hour, or 2 hours of culture. Data are representative of 3 independent experiments. (B) RPA analysis shows mRNA levels of Bcl-xL, Bak, and Bax of a representative experiment (left), and bars represent quantitative analysis of Bcl-xL (Bcl-xL/L32 mRNA ratio), Bax (Bax/L32 mRNA ratio), or Bak (Bak/L32 mRNA ratio) mRNA levels in WT (□) and TG (▦) uncultured thymocytes (0 h) or from thymocytes cultured for 1 or 2 hours from 3 independent experiments. (C) Apoptosis of WT (□) or TG(▦) thymocytes cultured for 24 or 48 hours. Bars represent the mean values ± 1 SEM from 10 independent experiments. ^*P < .05.