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

. 2018 Jul 18;8:10888. doi: 10.1038/s41598-018-29239-3

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

© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

EHMT2 promote methylation of MDC1 at lysine 45. (a) Schematic illustration of the full-length (FL) and deletion mutant (N1) of MDC1. Asterisk indicates the position of lysine 45 (wild type; WT or the alanine-substituted mutant; K45A). FHA, forkhead-associated domain; SDT, Ser-Asp-Thr repeats; TQXF, Thr-Gln-X-Phe repeats; PST, Pro-Ser-Thr repeats; BRCT, tandem BRCT domain. (b–d) Immunoblotting analysis of whole-cell extracts from U2OS cells transfected with (b) HA-tagged MDC1 (WT or K45A) and control or MDC1-targeting siRNAs, (c) HA-tagged MDC1-N1 (WT or K45A), and (d) HA-tagged MDC1-N1 (WT) and control, two EHMT1-targeting siRNAs (EHMT1 siRNA-1 and siRNA-2) or three EHMT2-targeting siRNAs (EHMT1 siRNA-1 siRNA-2 and siRNA-3; see methods), using the indicated antibodies. Quantification of methylation levels in histone H3 and MDC1 shown in right panels. Relative density was calculated based on signal intensities normalized against levels of total histone H3 (upper panel) and total MDC1-N1 (lower panel). Error bars, SD from triplicate measurements. (e) Immunofluorescence analysis of U2OS cells transfected with HA-tagged MDC1 (WT or K45A) and MDC1-targeting siRNA, and co-immunostained with indicated antibodies at 2 h after exposure to neocarzinostatin (NCS, 50 ng/ml for 15 min). A representative image of each treated or control cells is shown, as indicated. Scale bar, 10 μm.