Figure 6.
Analysis of Plk1 localization to mitotic structures in cells depleted of dynactin subunits. (A) Plk1 fluorescence intensities in cytokinetic bridges (B) and at spindle poles (C) of cells co-stained for tubulin (MT) were quantified, normalized to microtubule pixel values in the same area and expressed as per cent of controls (mean±s.e.m., n>150 cells). The loss of midbody density seen in (B) is commonly seen in cells depleted of dynactin subunits. Bar=5 μm. (D through K) Kinetochore localization of Plk1 and 3F3/2. (D, E) Representative images from asynchronous Cos7 populations fixed and stained for Plk1 (green, left panels) or 3F3/2 (green, right panels) plus anti-centromere antigen (ACA; red). Representative prometaphase cells are shown here. (E) Cells stained after brief nocodazole treatment are shown. Bar=5 μm. The insets (3 ×) show merged images of Plk1 or 3F3/2 with ACA. (F through K) Quantitative analysis of Plk1 (F, H, J) and 3F3/2 (G, I, K) fluorescence intensity at kinetochores, normalized to ACA pixel values (mean±s.e.m., n>600 kinetochores, 15 cells). (F, G) Cells transfected with different dynactin subunit siRNAs in the presence (+) or absence (−) of the wt p27 rescue vector. (H, I) Cells stained after brief nocodazole treatment. (J, K) Cells cotransfected with control or p27 siRNAs and the rescue vectors indicated (vector=pCAGIG lacking an insert).
Source data for this figure is available on the online supplementary information page.
