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Fig. 8. Myc-Ect2_1-333 mislocalizes to a ring-like structure at the cell cortex near the midbody. (A) The indicated myc-Ect2 constructs were overexpressed for 48 hours in HeLa S3 cells before fixation with paraformaldehyde and permeabilization with Triton X-100. Cells were then stained with anti-myc 9E10 in red (left) and anti--tubulin antibodies in green (middle). Merged images including DNA stained with DAPI (blue) are shown on the right. (B) Higher resolution, deconvolved images of the Ect2_1-333 expressing cells. The three times enlargements on the right show only the myc-Ect2 staining. (C) The myc-Ect2_1-333 stable cell line was induced for 36 hours and stained with anti-MKlp1 and anti-MgcRacGAP antibodies (middle) in red, anti--tubulin antibodies (left) in green and DNA was labeled with DAPI in blue. Merged images are shown on the right. (D) Quantitative analysis of the number of cells showing displaced myc-Ect2 fragments around the midbody (as shown in A and B). Histogram shows the results of three independent experiments (120 cells each) and bars indicate s.d. Bars, 10 µm.

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