Figure 1

Induction of mitotic accumulation by RSK2 depletion. (a) Representative images showing mitotic accumulation at indicated time points after each siRNA transfection (left panel). Cells stained with aceto-orcein were scored for mitosis at indicated time points after each siRNA transfection (right panel, n=300). Results are shown as the mean±s.d. from three independent experiments; *P<0.05, **P<0.01 by Student’s t-test. Cell lysates from the indicated samples were subjected to western blot analysis using indicated antibodies.(b) Mitotic index was scored after transfection of siRNA targeting different sequences of RSK2 (siRSK2-1; n=300). Results are shown as the mean ±s.d. from three independent experiments; **P<0.01 by Student’s t-test. (c) Transfection of each RSK2 plasmid DNA into cells was performed 24 h after siRNA transfection. Mitotic cells were scored 48 h after the RSK2 plasmid DNA transfection. (R2, siRSK2; Vec, vector DNA; WT-R2, wild-type RSK2; K100A, kinase-dead RSK2.) Results are shown as the mean±s.d. from three independent experiments (n=300); **P<0.01, ***P<0.001 by Student’s t-test. (d) Mitotic index was, respectively, scored after BI-D1570 treatment. Results are shown as the mean±s.d. from three independent experiments (n=300). *P<0.05, ***P<0.001 by Student’s t-test. siRNA, short interfering RNA.