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. Author manuscript; available in PMC: 2011 Jun 1.
Published in final edited form as: Eur J Cancer. 2010 Mar 18;46(9):1668–1678. doi: 10.1016/j.ejca.2010.02.017

Figure 3.

Figure 3

EM011 induces activation of mitotic checkpoint. A. Mitotic arrest is mediated by spindle-assembly checkpoint, as visualized microscopically by immunofluorescent staining of BubR1 (green) at 24 hr post-treatment. B. Immunoblot analysis also showed BubR1 accumulation until 36 hr of drug treatment. β-actin was used as a loading control. C. EM011 treatment for 72 hr induces apoptosis in PC-3 cells. Fragmented DNA in the terminal apoptosis stages was quantified using terminal deoxynucleotidyl transferase–mediated bromo-dUTP reaction (TUNEL assay) in EM011-treated cells. After the indicated incubation periods, cells were processed for a flow cytometry-based terminal deoxynucleotidyl transferase-mediated BrdUTP reaction. The addition of BrdUTP to the terminal deoxynucleotidyl transferase reaction labels DNA strand breaks that are then detected by an Alexa-Fluor 488-labeled anti-bromodeoxyuridine antibody. DNA content was determined using propidium iodide (PI, x axis). The number of apoptotic cells is indicated by the number of Alexa-Fluor 488-positive cells (vales on the top of the cytogram). Data from a representative experiment of two experiments.