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Figure 4 | Journal of Hematology & Oncology

Figure 4

From: Sodium selenite alters microtubule assembly and induces apoptosis in vitro and in vivo

Figure 4

Cyclin B1 interacted with CDK1 and induced the down-regulation of the anti-apoptotic protein Mcl-1. (A) CDK1 interacted with Cyclin B1 and Mcl-1. After cells were treated with selenite for 24 h, CDK1 was immunoprecipitated from the cell lysates using a CDK1 antibody, and the CDK1, Cyclin B1 and Mcl-1 in the precipitates were detected by western blotting. This experiment was repeated at least 3 times. (B) Inhibiting the expression of Cyclin B1 reversed the decrease of Mcl-1. Cells were transfected with siRNAs targeting Cyclin B1 and treated with 20 μM selenite for 24 h. Cyclin B1 and Mcl-1 were detected by western blotting. This experiment was repeated at least 3 times. (C) Inhibition of CDK1/Cyclin B1 reversed the down-regulation of Mcl-1. Roscovitine (10μM) was added to cells for 1 h before they were exposed to sodium selenite (20 μM) for 24 h. The changes of Mcl-1 detected by western blotting. This experiment was repeated at least 3 times. (D) The expression of Mcl-1 was inhibited by siRNA interference. Cells were transfected with siRNAs targeting Mcl-1 and treated with selenite (20 μM). Mcl-1 levels were determined by western blotting. This experiment was repeated at least 3 times. (E) Mcl-1 exhibited a protective role in HL60 cells. After Mcl-1 was knocked down, cell viability was assessed using the CCK-8 kits. The experiment was repeated three times, the mean value was determined, and *P < 0.05, when compared to untreated cells. (F) Inhibiting the expression of Mcl-1 increased the apoptotic ratio of the cells. Cells were transfected with siRNAs targeting Mcl-1 and treated with selenite (20 μM) for 24 h. The apoptotic ratio was determined by flow cytometry after the cells were stained with Annexin-V and PI. This experiment was repeated at least 3 times.

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