FIG. 6.

Reduced G₂/M inhibition by M3 and increased G₂/M inhibition by M1. (A) Asynchronous U2-OS cells were transiently transfected with vector (V), wild-type (wt) Wee1 (with two different amounts of plasmid [μg]), M1, and M3; harvested at 40 h; and subjected to flow cytometry (x axis, DNA content; y axis, relative cell number; profiles normalized to the highest peak) or immunoblotting for exogenous Wee1 (Myc tag). The percentages of cells in each cell cycle phase are listed. Note that the M1 G₂/M fraction was nearly as high as that of the wild type expressed at substantially higher levels, and the M1 S phase fraction was lower. (B) Cells transfected with the wild type, M1, or M3 were synchronized by mimosine, released for the designated times, and subjected to flow cytometry.