Figure 5.

Inhibition of cytochrome c release and apoptosis by Akt, but not by Bcl-2 or Bcl-xL, is dependent on glucose. Rat1a, Rat1a/MyrAkt, Rat1a/Bcl-2, and Rat1a/Bcl-xL cells were grown in glucose-free Dulbecco's modified Eagle medium with 1 mM pyruvate. Apoptosis was induced by growth factor withdrawal/UV (50 J/m²), and at the indicated times cells were fixed and stained with anti-cytochrome c antibody or DAPI. Three independent experiments were scored and the results are shown as the mean (±SE). (A) Analysis of cytochrome c release 0 h and 3.5 h after apoptosis induction in the absence of glucose and in untreated control cells (P < 0.002). (B) Percentage of apoptotic cells as measured by DAPI staining 0 h and 5 h after induction of apoptosis in the absence of glucose (P < 0.001). (C) Apoptosis as measured by DAPI staining in the presence of 20mM 5-thioglucose or 20mM 2-deoxy-glucose 5 h after induction of apoptosis (P < 0.002).