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

. 2019 Apr 16;10:568. doi: 10.3389/fimmu.2019.00568

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

Copyright © 2019 Eskandarian, Fliegauf, Bulashevska, Proietti, Hague, Smulski, Schubert, Warnatz and Grimbacher.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

The results of confocal microscopy with epitope-tagged constructs were consistent with the results of Immunofluorescent microscopy. (A) NIH3T3 cells, transiently overexpressing epitope-myc/FLAG-tagged wildtype IKAROS and myc-tagged mutant forms were incubated with monoclonal rabbit anti myc-tag-antibody and mouse monoclonal anti-FLAG M2 antibody and stained with goat anti-rabbit Alexa fluor 488 (green) and goat anti-mouse Alexa 546 (red). In NIH3T3 cells transfected with epitope-myc/FLAG-tagged wildtype IKAROS the speckled nuclear localization was observed in contrast to a diffuse nuclear staining, observed in NIH3T3 cells expressing the myc-tagged Arg143Trp, Cys150Arg, Arg162Gln, and Lys286* mutant forms. (B) Wildtype components were dominant to form IKAROS complexes in PC-HC sites. NIH3T3 cells were co-transfected with 50% of myc/FLAG-tagged wildtype IKAROS and 50% of myc-tagged, expressing each mutant form. The diffuse nuclear localization of the mutant forms was significantly reduced in NIH3T3 cells co-transfected with equal amounts of the FLAG-tagged wildtype IKAROS together with the myc-tagged Arg143Trp, Cys150Arg, and Arg162Gln. Accumulation of FLAG-tagged wildtype proteins, observed as a punctuated nuclear staining pattern in the co-transfected samples, showed the dominance of wildtype components to form IKAROS complexes in PC-HC sites.