Fig. 5. Reduced 14-3-3 binding to Wee1 during mitosis. HeLa cells were transfected with vector alone (A, Lane 1) or with vectors encoding myc-Wee1 (A, Lanes 2 and 3), myc-p49Wee1 (B, Lanes 1 and 2), myc-p49Wee1(K328R) (B, Lanes 3 and 4), or myc-p49Wee1(S642A) (B, Lane 5). Five h after transfection, nocodazole was added to the indicated cultures. Cells were harvested and lysed 16 h after nocodazole addition. Lysates were incubated with anti-c-Myc (9E10) agarose, and precipitates were resolved on a 12% gel. Wee1 and 14-3-3 were detected by immunoblotting as described in Fig. 4 .