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

. 2020 Nov 6;11:5647. doi: 10.1038/s41467-020-19159-0

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

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2020

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

Fig. 6 — a Schematic representation of ICF4 associated human LSH mutations (red box) and alignment with murine LSH. b Western blot analysis to confirm the expression of three LSH-FRB-V5 mutant proteins and ZnF-FKBP-HA protein in ICF4 associated LSH CIP system compared to that in wild-type LSH CIP system using V5 and HA antibodies. Cells lacking LSH-FRB-V5 fusion protein expression served as controls. c ChIP-qPCR analysis to determine the recruitment of ICF4 associated LSH mutants and wild-type LSH to the engineered Oct4 locus in the CIP system after rapamycin treatment at indicated time points using V5 antibody (n = 3 independent experiments). Cells lacking LSH fusion protein expression served as controls. Data are represented as mean ± SD. d Flow cytometry was used to measure reporter-GFP expression in ICF4 derived LSH mutants CIP system compared to that in wild-type or ATP mutant LSH (K 237 A) CIP system after 0 (red) and 5 (blue) days treatment of rapamycin. e ChIP-qPCR analysis to assess the dynamic changes of macroH2A1, macroH2A2 and H2A enrichment at the CiA: Oct4 locus in the engineered mouse ES cells tethered with ICF4 associated LSH mutants or wild-type LSH with rapamycin treatment for 0 and 5 days (n = 3 independent experiments). Data are represented as mean ± SD. Significance assessed using one-way ANOVA with Tukey’s multiple comparison test (wild type: *adjusted p < 0.05, ** adjusted p < 0.01 and ***adjusted p < 0.001; Q 682 R: ^❖adjusted p < 0.05 and ^❖❖adjusted p < 0.01; L 784 del: ^#adjusted p < 0.05). f Flag-IP of indicated proteins confirmed by western blot analysis in U2OS cells with a stable expression of wild-type Flag-LSH or ICF4 associated Flag-LSH mutants. Cells without a stably integrated of Flag-LSH expression vector served as controls. Source data are provided as a Source Data file.