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

. 2017 Apr 3;7:45022. doi: 10.1038/srep45022

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

Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

PMC Copyright notice

Chromatin association of CFDP1 in HeLa cells (A) Western blot analysis of fractionated HeLa cells after synchronization in interphase (G1/S) or metaphase (see material and methods). As metaphase synchronization controls, the histone 3 phosphorylated in serine 10 (H3p) and SMC2 condensin have been used. H3p is a DNA condensation marker which is found only in mitotic chromatin, while SMC2 increases in condensed chromatin. As expected, H3p is specifically present in the chromatin-bound fraction (P3) of HeLa cells synchronized in metaphase. In the same fraction, the amount of SMC2 is higher compared to that of cells synchronized in G1/S. CFDP1 is detected at comparable amount in the chromatin-bound fractions (P3) of both interphase and metaphase, similar to histone H3. (B) Schematic representation of Flag-CFDP1 tagged proteins; Flag-CFDP1 full-length, isoform 1(aa 1-299); Flag-CFDP1-Nt, isoform 2 (1–217) and Flag-CFDP1-Ct, BCNT domain (218–299). (C) From left to right panels: DAPI (blue), α-tubulin (green), Flag-CFDP1 (red) and merge. (D) From left to right panels: DAPI (blue), α-tubulin (green), Flag-CFDP1-Nt (red) and merge. (E) From left to right panels: DAPI (blue), α-tubulin (green), Flag-CFDP1-Ct (red) and merge. (F) Western blot analysis of fractionated HeLa cells after transfection with Flag-CFDP1 or truncated variants. Flag-CFDP1 is detected in soluble (S2 and S3) and chromatin-bound (P3) fractions, while Flag-CFDP1-Nt and Flag-CFDP1-Ct are found only in the soluble fractions (S2 and S3). MEK2 and H3 are specific controls for soluble and chromatin fractions, respectively. Anti-MEK2, mitogen-activated protein (MAP)/ERK kinase 2, was used as cytoplasmic contamination control in P3 fraction; anti-histone H3 was used to monitor chromatin contamination in soluble fraction. WCE = whole cell extract; S2 = cytoplasmic soluble fraction; S3 = nuclear soluble fraction, P3 = chromatin-bound fraction.