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. 2014 Oct 9;10(10):e1004672. doi: 10.1371/journal.pgen.1004672

Figure 5. Plk1 regulates substrate cleavage of Scc1 but neither the activity of separase nor the cleavage of kendrin.

Figure 5

(A) The level of cyclin B1 and cleaved kendrin do not change upon Plk1 inhibition. FL indicates the non-cleaved kendrin and CL the cleaved kendrin product. The asterisk indicates protein bands that are unspecifically recognized by the anti-kendrin antibody. (B) mCherry-Kendrin(2059-2398)-eGFP-PACT stably expressing HeLa cells were analyzed for the cleavage of the sensor upon Plk1 inhibition. Experiment was performed as described in Figure 4A. The two-fold magnifications on the bottom right of the figures highlights the centrosomes. The scale bar represents 10 µm. (C) The quantification of (B). mCherry/eGFP ratio at centrosomes was quantified just after (t = 0) and 48 min after (t = 48) addition of the Plk1 inhibitor BI2536. Data were normalized to the average mCherry to eGFP ratio at t = 0 min. (D) The levels of γ-tubulin and Espl1 signals at the centrosome were plotted using Whisker-Box plot after Plk1 (BI2536) inhibition. The level of Espl1 did not change upon Plk1 inhibition although the γ-tubulin signal declined compared to nocodazole treatment as a control to maintain the cells at prometaphase. Statistical analysis was performed with one-way ANOVA (*** represents p<0.001). N≥30, Error bars: SD.