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. 2016 Feb 3;7:10597. doi: 10.1038/ncomms10597

Figure 7. NuSAP governs Kid-generated PEF during chromosome oscillation.

Figure 7

(a) Representative images and colour project of MT gliding assay with Kid or NuSAP and Kid proteins. Arrows represent one MT gliding at different time points. Images were acquired in a 2 s interval for 5 min. Scale bar, 3 μm. (b) Kymographs showing a sample microtubule gliding on Kid-coated surfaces in the presence or absence of NuSAP protein. (c) Bar chart representing the average velocity of MT gliding on Kid-coated surfaces in the presence or absence of NuSAP protein. Error bars represent±s.d. *P<0.01, by t-test. (d) Bar chart representing the average of the amplitude (black) and centre velocity (grey) of centromere oscillation in control- or NuSAP/Kid-double-depleted metaphase cells. Error bars represent +s.e.m. *P<0.001, by t-test. (e) Bar chart representing the average of the amplitude (black) and centre velocity (grey) of centromere oscillation in control- or NuSAP/Kid-double-depleted monopolar cells. Error bars represent +s.e.m. *P<0.001, by t-test. (f) The kinetochore MTs in metaphase HeLa cells transfected with control siRNA, NuSAP siRNA, GFP-vector or GFP-NuSAP after cold treatment. Cells were stained with an anti-α-tubulin antibody and DNA with Hoechst 33342. Scale bar, 5 μm. (g) Representative images of PAGFP α-tubulin stability in metaphase HeLa cells transfected with control siRNA, NuSAP siRNA, mCherry vector or mCherry-NuSAP after photoactivation. Dotted squares represent photoactivated region. Scale bar, 5 μm. (h) Bar chart of the half-lives of spindle MTs calculated with a double exponential regression fitting of photoactivation assays. n (control siRNA)=14 cells, n (NuSAP siRNA)=13, n (mCherry)=13, n (mCherry-NuSAP)=12. Error bars represent ±s.d. (i) FRAP assays; representative images of MT dynamics in metaphase stable mCherry-α-tubulin HeLa cells transfected with control siRNA, NuSAP siRNA, GFP-vector or GFP-NuSAP after photobleaching. Dotted squares represent the photobleaching region. Scale bar, 5 μm. (j) Dot plot representing the half-life of kinetochore microtubules (slow phase) and interpolar microtubules (fast phase) calculated according to a double exponential regression curve with FRAP assays in control siRNA-, NuSAP siRNA-, GFP-vector- or GFP-NuSAP-transfected metaphase cells. Mean±s.e.m. are indicated.