WT4 cells activate the anaphase-promoting complex. (A) Sequencing of the p53 hot spot in early passage and immortalized WT4 cells, compared with wild-type p53. (B) p53 Luciferase (pGL13) activity between wild-type p53 (FL) and WT4-p53 in Jurkat cells. (C) Real-time PCR expression of p21^WAF and mdm2 in PBMCs, C8166, and WT4 cells after gamma irradiation (10 Gy). RNA was extracted 2.5 hours after irradiation. Real-time PCR was performed in duplicate; error bars represent the C_T SD of the gene of interest. (D) Western blot analysis of cyclin A, cyclin B, Skp2, p27^KIP, and p21^WAF in WT4 and MT2 cells compared with nontransduced resting (RPBMCs) or activated (ActPBMCs) PBMCs. (E) Mitotic spindle checkpoint studies. Cells were treated for 24 hours with either nocodazole (400 ng/mL) or paclitaxel (10μM), followed by cell-cycle analysis by flow cytometry.