Figure 5.

Inhibition of Akt activity by citrate confers the induction of cyclin B1–p-CDK1 (Thr 161) complexes and G₂/M phase arrest. At 12 h after transfection with an empty vector, HA-CA Akt1, or HA-DN Akt1, the cells were treated with either vehicle (−) or citrate (10 mM) for 36 h. (A,B) The levels of the indicated proteins in the cell lysates and cell cycle phase were determined using Western blot analysis with specific antibodies and flow cytometric analysis of PI-stained cells, respectively. The values are presented as the means ± standard error of three independent experiments. * p < 0.05: significantly different from empty vector-transfected vehicle-treated cells or empty vector-transfected citrate-treated cells. (C) The antibody used for coimmunoprecipitation is indicated at the top. The proteins from the immunoprecipitated complexes were detected using Western blotting with specific antibodies. Normal IgG was used as a control for antibody specificity.