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

. 2012 Mar 1;287(17):13674–13685. doi: 10.1074/jbc.M111.320705

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

© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

PMC Copyright notice

FIGURE 4. — Sprouty2 and Gα_o compete for GRIN2 binding. A, the region of 200–300 aa of GRIN is essential for Sprouty2 and Gα_o binding. Left panel, HEK-293T cells were co-transfected with the indicated expression constructs, and total cell lysates were prepared as described previously. Lysates were immunoprecipitated (IP) with the anti-Gα_o antibody or with the M2-FLAG antibody and immunoblotted (WB) with the anti-HA antibody (panel 1), with the anti-Gα_o antibody (panel 2), and with the M2-FLAG antibody (panel 3). Panel 3 contains bands for Gα_o (single band) and FLAG-Sprouty2 (doublet bands). Right panel, whole cell lysates (WCL) were immunoblotted with the anti-HA antibody (panel 1), with the anti-Gα_o antibody (panel 2), and with the M2-FLAG antibody (panel 3). Panel 3 contains bands for Gα_o (single band) and FLAG-Sprouty2 (doublet bands). Representative immunoblots from one of two independent experiments are shown. mSp2, mouse Sprouty2; mG2, mouse GRIN2. B, Sprouty2 and Gα_o compete for GRIN2 binding. HEK-293T cells were co-transfected with the indicated plasmids: Gα_o Q205L (Q/L) mutant (1.5, 3.0, or 6.0 μg), FLAG-mSprouty2 (2.0, 4.0, or 6.0 μg), and HA-mGRIN2 (4 μg). Vector pcDNA3.1 was co-transfected to make the total amount of DNA transfected equal. Lysates were prepared as described previously and immunoprecipitated with anti-HA antibody. Immunoprecipitates were immunoblotted with anti-Gα_o (panel 1), M2-FLAG (panel 2), or anti-HA (panel 3) specific antibodies. The triangle indicates increasing concentration of DNA transfected, and the rectangle indicates the same concentration of DNA transfected. A representative blot from three independent experiments is shown. Lower panel, whole cell lysates were immunoblotted with anti-Gα_o (panel 1), M2-FLAG (panel 2), or anti-HA (panel 3) specific antibodies. Representative immunoblots from one of three independent experiments are shown. mSpry2; mouse Sprouty2. C, corresponding densitometry for the average of 3–6 independent experiments testing the following DAN concentrations: Gα_o Q205L (Q/L) mutant (1.5, 3.0, 5.0, or 6.0 μg), FLAG-mSprouty2 (2.0, 4.0, 5.0 or 6.0 μg), and HA-mGRIN2 (4 μg). AU, arbitrary units. Error bars are S.E.