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Fig. 5. Generation of ROS following exposure to menadione or chemical hypoxia (CoCl₂ treatment). (A) Embryoid bodies were treated for 24 hours with 20 µM menadione. Subsequently, the cell culture medium was exchanged with medium devoid of menadione, and ROS were monitored at different times after removal of menadione. (B) ROS generation in the presence of either menadione (20 µM) or CoCl₂ (50 µM). Determination was performed immediately after exposure. The upregulation of CT-1 by menadione (C) and CoCl₂ (D) was inhibited by either the free radical scavenger vitamin E (20 µM) or NMPG (20 µM). Embryoid bodies were treated at day 4 of cell culture with either menadione (20 µM) or CoCl₂ (50 µM) in the presence of free radical scavengers. After 24 hours CT-1 protein expression was determined by semiquantitative immunohistochemistry. ^*P<0.05, significantly different to levels in the relative controls. ^#P<0.05, significantly different to levels in the CoCl₂ or menadione-treated samples before addition of scavengers.

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