Fig. 2.

Subcellular localization of VRK1 and AURKB in mitosis. VRK1 and AURKB localizations during cell cycle progression and mitosis. 24 h after plate the cells, U2OS cells were treated with serum-free medium for 72 h, to arrest the cells at G0/G1, or with double-thymidine block to arrest cell cycle at S-phase, or with double-thymidine followed nocodazole treatment to arrest cells at G2/early mitosis, or after double-thymidine and nocodazole treatment, released from the arrest during 360 min. The known AURKB distribution in mitosis is also used as an internal control. In immunofluorescence, AURKB was detected with rabbit monoclonal anti-AURKB (N-term) antibody. Human VRK1 was detected using mouse monoclonal anti-VRK1 antibody. The flow cytometry profile of synchronized cells and their release is shown in Fig. S1. A more detailed image with additional time points in the thymidine/nocodazole release is shown in Supplementary Fig. S2. Immunofluorescence experiments were performed three times