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. 1998 Jun 9;95(12):6785–6790. doi: 10.1073/pnas.95.12.6785

Figure 5.

Figure 5

(A) Changes in Tyr-15 phosphorylation of cdc2 and cdk2 during Fas- and staurosporine-induced apoptosis. The same filter was blotted with anti-cdc2, anti-cdk2, and a phospho-specific cdc2 (cdk2) antibody against Tyr-15. The cell death percentage was measured by MTT assay (20). (B) Cdc25 activity during Fas-induced apoptosis. Samples from control extract (−α Fas), apoptotic extract (+α Fas), and nocodazole arrested extract (N-arrested, ≈50% in mitosis) were analyzed with anti-cdc25 immunoblotting (Upper). The relative cdc25 activities are shown graphically (Lower). (C) Anti-Wee1 blot of the extracts. (D) Wee1 activity during apoptosis. (Middle and Bottom) Western blot analysis of the active cyclin B/cdc2 beads treated with different extracts. (Top) H1 kinase activity of the same corresponding beads. Each extract (≈5 mg/ml total proteins) made in Nonidet P-40 lysis buffer was analyzed at three different dilutions: 1, 1/5, and 1/25. Five milligrams per milliliters BSA was used as a negative control.