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. 2016 May 23;113(23):6490–6495. doi: 10.1073/pnas.1523157113

Fig. 5.

Fig. 5.

RUNX3 is involved in regulation of mitosis. (A) U2OS cells were treated with siRNA to deplete AURKA or AURKB proteins. Nocodazole-treated cells (Noc) were harvested by mitotic shake-off. RUNX3 and Aurora proteins were detected by immunoblot. (B) Densitometric analysis of RUNX3 expression after AURKA or AURKB knockdown as in A; three independent experiments were done. (C) U2OS cells were treated with nocodazole before addition of MG132 and Aurora kinase inhibitors (MLN8405 and ZM4473439) for 4 h. (D) Cells were treated with nontargeting siRNA (siControl) or RUNX3 siRNA (siRUNX3). Nocodazole was added for the last 16 h. Immunoblot was done with the indicated antibodies. (E) Immunoblot of U2OS cells treated with siRNA targeting RUNX1, RUNX2, and RUNX3. R123 indicates treatment with all three RUNX siRNA. (F) HeLa.Fucci cells were treated with RUNX3 siRNA and subjected to double thymidine block. Cells were monitored by time-lapse microscopy following release from block. Metaphase cells (visualized as mitotic rounding up) were scored. Numbers of cells counted for siControl and siRUNX3 are 447 and 492, respectively. The data represented mean ± SEM of three independent experiments. *P = 0.03, **P = 0.005 (Student’s t test).