Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2011 Jan 28;6(1):e16307. doi: 10.1371/journal.pone.0016307

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Mattiuzzo et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

PMC Copyright notice

(A) Western blot analysis using an Hec1 antibody before (-) and after (+) doxycycline addition in HeLa clones expressing Hec1 (D clones), Hec1-EGFP (C clones) or EGFP-Hec1 (A clones). (B) Densitometric analysis of Hec1 expression in the different clones as measured by the ratio of total Hec1 (endogenous + exogenous) in induced (doxy) vs uninduced (cnt) conditions after normalization for vinculin expression. Data are mean ± SE of the ratios obtained in clones expressing untagged Hec1 (n = 10), Hec1-EGFP (n = 8) or EGFP-Hec1 (n = 8). (C) EGFP-Hec1 (green) localization after doxy induction in a prometaphase cell stained for CREST (blue) and tubulin (red). The boxed 2-fold enlargement shows EGFP-Hec1 localization with respect to the inner kinetochore marker CREST. Bar, 5 µm. (D) Chromosome congression and spindle organization before (-doxy) and after (+doxy) induction of Hec1-EGFP or EGFP-Hec1 expression as shown by α-tubulin (red) and DAPI (blue) staining. In uninduced samples Hec1 is visualized by antibody staining (green). Hec1-EGFP/-doxy: normal metaphase; Hec1-EGFP/+doxy: late prometaphase with unaligned chromosomes; EGFP-Hec1/-doxy: normal prometaphase; EGFP-Hec1/+doxy (fourth row): disorganized bipolar prometaphase; EGFP-Hec1/+doxy (last row): disorganized multipolar prometaphase. Bar, 5 µm. (E) Quantitative analysis of prometaphase abnormalities in Hec1-EGFP or EGFP-Hec1 expressing clones. Data are mean ± SE of 300 prometaphases (PM) for each condition in three independent experiments. Alignment defects were significantly higher in Hec1-EGFP expressing mitoses than in uninduced cells (P<0.01, t-test); disorganized bipolar prometaphases (P<0.05, t-test) and disorganized multipolar prometaphases (P<0.01, t-test) were significantly higher in EGFP-Hec1 expressing mitoses than in uninduced mitoses.