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

. 2016 Feb 1;27(3):434–441. doi: 10.1091/mbc.E15-07-0546

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

© 2016 Fisher et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

“ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

PMC Copyright notice

FIGURE 2: — Et/Lat can bind to intracellular JAK/STAT components. (A) Visualization of Dome and Et/Lat protein domain structure. Clustal sequence alignment for intracellular portion of Dome and Et/Lat. Transmembrane domain (TM; black bar), putative binding sites for Hop (HBS), and STAT92E (SBS) are highlighted. Tyrosine residues in Et/Lat are shown with bold letters. SP, signal peptide; CBM, cytokine-binding motif; FNIII, fibronectin 3 domains; TM, transmembrane domain; HBS, Hop-binding site; SBS, STAT binding site; aa, amino acids). Percentage identity between the indicated domains. (B) Dome-FLAG can coimmunoprecipitate Hop-HA, but this is greatly reduced when the putative Hop-binding site is mutated (Dome P925I). Tyrosine phosphorylation (pTyr) of Dome is also reduced in the Dome-P925I. (C) STAT92E transcriptional reporter activity assay. JAK/STAT pathway signaling is greatly reduced in cells expressing Dome P925I even in the presence of Upd2 ligand, **p < 0.01. (D) Immunoprecipitation assays show that both Dome-FLAG and Et/Lat-FLAG can coprecipitate Hop-HA. (E) Immunoprecipitation assays after transfection of the indicated Dome-FLAG and Et/Lat-FLAG proteins show that both proteins can coprecipitate endogenous STAT92E. dsRNA targeting the labeled mRNAs was added as indicated; ctrl, a nontargeting dsRNA. Knockdown of endogenous Dome (final lane) did not disrupt the ability of Et/Lat to bind to STAT92E. Upd2-conditioned medium was added for 15 min before protein harvesting. EV, empty vector. (F) Immunoprecipitation of endogenous STAT92E from cells expressing FLAG-tagged Dome or Et/Lat shows that both receptors are coprecipitated. Treatment with dsRNAs targeting the endogenous receptors suggests that interactions detected are independent of potential heterodimer formation.