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

. 2014 Aug 15;28(16):1815–1826. doi: 10.1101/gad.243113.114

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

© 2014 Deyter and Biggins; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

PMC Copyright notice

Figure 1. — The FACT complex facilitates CENP-A^Cse4 degradation in vivo. (A) spt16-aid (SBY11265) cells were treated with vehicle (95% ethanol [−NAA]) or auxin (+NAA) at time 0, and lysates were analyzed for Spt16-AID levels using α-Spt16 antibodies at the indicated time points. Pgk1 is shown as a loading control. (B) spt16-aid (SBY11268) cells containing pGAL-3Flag-CSE4 were treated with vehicle (−NAA) or auxin (+NAA) to degrade Spt16 for 1 h, followed by a 1-h galactose induction of 3Flag-Cse4. Protein synthesis was inhibited by cycloheximide (CHX) addition at time 0, and lysates were monitored for 3Flag-Cse4 levels with α-Flag antibodies at the indicated time points. Pgk1 is a loading control. Quantification of the stability is in Supplemental Figure S1. (C) Fivefold serial dilutions of the indicated strains (SBY3, SBY8851, SBY8336, SBY8903, SBY6006, and SBY11186) were plated on glucose or galactose medium at 30°C. (D) Fivefold dilutions of the indicated strains (SBY3, SBY8851, SBY8336, SBY8903, SBY11295, and SBY11296) were plated on glucose or galactose medium at 23°C. (E) pGAL-3Flag-CSE4 was overexpressed with galactose in either spt16-aid (SBY11268) cells treated with (+NAA) or without (−NAA) auxin as in B or psh1∆ (SBY8903) cells. This was followed by cycloheximide addition for 30 min, and then whole-cell extracts (WCE) were fractionated into soluble and chromatin fractions. Cse4 levels were monitored in each fraction with α-Flag antibodies. Pgk1 and H2B are markers of the soluble and chromatin fractions, respectively.