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

. 2016 May 23;213(4):415–424. doi: 10.1083/jcb.201512034

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

© 2016 Liu and Mao

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 1. — The formin mDia2 is required to maintain CENP-A levels at centromeres. (A) Depletion of mDia2 does not affect CENP-A protein level. Immunoblotting analysis of HeLa cell lysates 48 h after transfection with control (GAPDH) and mDia2 siRNAs. (B) Immunofluorescence detection of CENP-A and CENP-B 48 h after transfection with the indicated siRNAs and the full-length mEmerald-tagged mDia2 (FL-mDia2) expression vector (DNA - DAPI). Transfected cells were identified by cotransfected fluorescence markers. Bar, 10 µm. (C) Scatterplot showing the distribution of normalized CENP-A integrated intensity per nucleus (mean ± SD overlaid with scatterplot) with the INCA method (Fig. S1). Control: n = 292; HJURP siRNA: n = 196; mDia2 siRNA: n = 260; mDia2 siRNA + FL-mDia2: n = 136; and mDia3 siRNA: n = 117 from three independent experiments. The p-value was computed using a two-tailed t test. (D) Depletion of mDia2 does not affect centromere numbers. Whisker-Tukey boxplots show the number of CENP-B foci. The boxes span the 25th to 75th percentile of the data, whereas the center bar denotes the median and + marks the mean. Control: n = 168, and mDia2 siRNA: n = 125, from three independent experiments. The p-value was computed using a two-tailed t test. (E) Depletion of mDia2 results in reduced levels of YFP-CENP-A at centromeres. Immunofluorescence images showing YFP-CENP-A in HeLa cells stably expressing YFP-CENP-A 48 h after transfection with indicated siRNAs (DNA - DAPI). Bar, 10 µm. (F) Quantification showing the mean levels of normalized integrated YFP-CENP-A intensity per nucleus (means ± 95% confidence intervals). Control (GAPDH siRNA): n = 456; HJURP siRNA: n = 250; and mDia2 siRNA: n = 158 cells from three independent experiments. The p-value was computed using a two-tailed t test.