Figure 2.

DEK protein is sharply reduced in mitotic cells. NIKS were treated with mimosine or serum starvation to induce cell cycle arrest in the G1 phase, with a combination of thymidine and aphidicolin to induce cell cycle arrest in S phase, and with nocodazole or underwent a mitotic shake off to enrich for cells in G2/M phase. Mitotic shake off occurred after treatment with colcemid 20 nM for 24 hours. (A) Cellular arrest in the expected phase of the cell cycle was verified by flow cytometry. Cells were pulsed with EdU for 2 hours then stained with 7AAD to determine DNA content. (B) Percent cells from the gated areas of G1, S, and G2/M were quantified from (A). (C) Cells were treated as in (A) and subjected to protein gel blot analysis to detect DEK protein (Aviva Systems Biology) levels throughout the cell cycle.