Fig. 7.

VRK1 alters the phosphorylation of H3 in Thr3 and the localization of AURKB in centrosomes. a VRK1 downregulation affects the phosphorylation of histone H3 on Thr3 residue. At the top are shown non-synchronized cells. At the bottom are shown U2OS cells in which siControl (siCt) or si-VRK1 (siV1-02) was performed, and 2 days later, cells were treated with nocodazole for 13 h. H3-T3ph was detected using a rabbit monoclonal anti-H3-T3ph and VRK1 was detected using a mouse monoclonal anti-VRK1 (1B5) antibody. b Depletion of VRK1 in nocodazole-treated cells caused a reduction in phopho-Rb and phosphorylation of H3 in Thr3 and Ser10. The relative values of H3 coprecipitating with AURKB are the mean of three experiments, and value 1 is the reference in unsynchronized cells. c VRK1 downregulation affects the phosphorylation of histone H3 on Ser10 residue. At the top are shown non-synchronized cells. At the bottom are shown U2OS cells in which siControl (siCt) or si-VRK1(siV1-02) was performed and 2 days later cells were treated with nocodazole for 13 h. H3-S10ph was detected using a rabbit monoclonal anti-H3S103ph and VRK1 was detected using a mouse monoclonal anti-VRK1 (1B5) antibody. d Depletion of VRK1 altered the localization of AURKB on centromeres in nocodazole-treated cells. The U2OS cells were treated during 13 h with nocodazole. Immunofluorescence experiments were independently performed three times