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

. 2011 Jan 13;286(10):8688–8696. doi: 10.1074/jbc.M110.141754

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

© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

PMC Copyright notice

FIGURE 1. — Identification of α-pPER2 cross-reacting species as rpS6. A, α-pPER2-662/665/668 reacts with overexpressed hPER2 and an endogenous 34-kDa protein. HEK 293T cells were transfected with either empty vector (EV) or hPER2. Cell extracts were analyzed by immunoblotting with α-pPER2-662/665/668 antibody. hPER2 is indicated by an arrow, and the endogenous 34-kDa protein is indicated by an asterisk. B, immunoprecipitation of 34-kDa protein. HEK 293T cell extracts were subjected to immunoprecipitation with mouse IgG or α-pPER2-662/665/668 antibody. Immunoprecipitates (IP) and whole cell lysates (WCL) were analyzed by immunoblotting with α-pPER2-662/665/668 antibody. C, summary of proteomic analysis of cross-reacting 34-kDa protein. Two replicate immunoprecipitated 34-kDa bands were excised from Coomassie Blue-stained SDS-PAGE gels and submitted to ProtTech Inc. for analysis by LC-MS/MS. A summary of the sequencing report is shown. D, α-pPER2-662/665/668 antibody reacts with rpS6. HEK 293T cells were transfected with empty vector, HA-rpS3, or HA-rpS6. Where indicated, cell extracts were treated with λ-phosphatase (PPase) prior to analysis by SDS-PAGE and immunoblotting with α-pPER2-662/665/668, α-rpS6, α-tubulin, α-FLAG, and α-HA antibodies. These data are representative of three independent experiments.