Effects of PD0166285 on the G2 phase. A: Immunoblot analysis of anti-cdc2, anti-phosphorylated cdc2-tyr15, and Wee1 was performed in lysates of with 0.5 μM PD0166285 treated for 0 h, 2 h, or 4 h. B: Subcellular location of cyclin B was observed. After 4 h of treatment with 0.5 μM PD0166285, cells were fixed and doubly stained for immunofluorescence with PI (red) and anti-cyclin B1 (green). Shown are typical merged images obtained by confocal microscopy. Bar: 10 μm.