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

. 2021 Aug 25;10:e65872. doi: 10.7554/eLife.65872

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

© 2021, Takahashi et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

PMC Copyright notice

Figure 4. — (A) Requirement of the occludin homology domain (OHD) in MLL-ELL-mediated transformation. Various MLL-ELL constructs carrying mutations in the ELL portion were examined for the transformation of myeloid progenitors, as in Figure 1A. NHD: N-terminal homology domain; ALF: AF4/LAF4/FMR2 homology domain; pSER: poly-serine; A9ID: AF9 interaction domain; CHD: C-terminal homology domain; EHD: EAF family homology domain; DLXLS: DLXLS motif; SDE: SDE motif. (B) OHD-dependent association with AF4. Immunoprecipitation (IP)-western blotting of the chromatin fraction of virus-packaging cells, transiently expressing various HA-tagged MTMT-ELL fusion constructs, was performed. Endogenous AF4 proteins co-purified with MTMTh-ELL proteins were visualized by an anti-AF4 antibody. (C) Sequential recruitment of AF4 and EAF1 by ELL. HEK293TL cells (Okuda et al., 2015), which harbor GAL4-responsive reporter, were transfected with various combinations of FLAG-tagged GAL4 fusion proteins, xAF4, and HA-tagged EAF1 (EAF1h), and were subjected to chromatin immunoprecipitation (ChIP)-qPCR analysis. A qPCR probe near the GAL4-responsive elements (UAS) was used. The ChIP signals were expressed as the percent input with error bars (mean ± SD of PCR triplicates). TATA: TATA box; LUC: luciferase. (D) Effects of overexpression of AF4 or EAF1 on ELL complex formation. IP-western blotting of the chromatin fraction of HEK293TL cells, transiently expressing various combinations of FLAG-tagged GAL4-ELL proteins, xAF4, and EAF1h, was performed. (E) A model of the sequential association between ELL, AF4 family proteins, and EAF family proteins.

Figure 4—figure supplement 1. — (A) Requirement of the occludin homology domain (OHD) in MLL-ELL-mediated transformation. Various MLL-ELL constructs carrying mutations in the ELL portion were examined for the transformation of myeloid progenitors, as in Figure 1A. NHD: N-terminal homology domain; ALF: AF4/LAF4/FMR2 homology domain; pSER: poly-serine; A9ID: AF9 interaction domain; CHD: C-terminal homology domain; EHD: EAF family homology domain; DLXLS: DLXLS motif; SDE: SDE motif. (B) OHD-dependent association with AF4. Immunoprecipitation (IP)-western blotting of the chromatin fraction of virus-packaging cells, transiently expressing various HA-tagged MTMT-ELL fusion constructs, was performed. Endogenous AF4 proteins co-purified with MTMTh-ELL proteins were visualized by an anti-AF4 antibody. (C) Sequential recruitment of AF4 and EAF1 by ELL. HEK293TL cells (Okuda et al., 2015), which harbor GAL4-responsive reporter, were transfected with various combinations of FLAG-tagged GAL4 fusion proteins, xAF4, and HA-tagged EAF1 (EAF1h), and were subjected to chromatin immunoprecipitation (ChIP)-qPCR analysis. A qPCR probe near the GAL4-responsive elements (UAS) was used. The ChIP signals were expressed as the percent input with error bars (mean ± SD of PCR triplicates). TATA: TATA box; LUC: luciferase. (D) Effects of overexpression of AF4 or EAF1 on ELL complex formation. IP-western blotting of the chromatin fraction of HEK293TL cells, transiently expressing various combinations of FLAG-tagged GAL4-ELL proteins, xAF4, and EAF1h, was performed. (E) A model of the sequential association between ELL, AF4 family proteins, and EAF family proteins.