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. 2023 Nov 15;15(22):5424. doi: 10.3390/cancers15225424

Figure 1.

Figure 1

The ASPP family of proteins is phosphorylated during mitotic arrest. (A,B) HeLa cells were treated with dimethyl sulfoxide (DMSO), nocodazole (Noco, 100 ng/mL), or taxol (100 nM) overnight. Total cell lysates were electrophoresed on Phos-tag (A) or regular (B) SDS polyacrylamide gels and probed with indicated antibodies. The increased Cyclin B1 level indicates cells in G2/M phase. (C) S2.013 and PANC-1 cells were treated as in (A). Total cell lysates were probed with the indicated antibodies. (D) HeLa cells were treated with DMSO or taxol, and total cell lysates were further incubated with (+) or without (−) lambda phosphatase (λ ppase) prior to western blotting. (E) HeLa cells were treated with taxol overnight and subjected to the indicated kinase inhibitors for 1.5 h. MG132 was added along with the kinase inhibitors to prevent mitotic exit. The lysates were harvested for western blotting. The uncropped bolts are shown in Supplementary Materials.