Supplementary Materials

Supplementary Materials for:

Large-Scale Discovery of ERK2 Substrates Identifies ERK-Mediated Transcriptional Regulation by ETV3

Scott M. Carlson, Candace R. Chouinard, Adam Labadorf, Carol J. Lam, Katrin Schmelzle, Ernest Fraenkel, Forest M. White*

*To whom correspondence should be addressed. E-mail: fwhite{at}

This PDF file includes:

  • Methods. Detailed protocol for ERK2 substrate labeling and identification.
  • References
  • Fig. S1. Additional sites phosphorylated by ERK2 on IRS2, CDC42EP1, and ETV3.
  • Fig. S2. MS/MS spectrum used to identify ETV3 as a direct ERK2 target and the matching spectrum of a synthetic peptide.
  • Fig. S3. HA-tagged ETV3 exhibits MEK-dependent gel shift, which is reduced in phosphosite mutants, and an anti-HA antibody captures both the phosphorylated and the nonphosphorylated protein in ChIP.
  • Fig. S4. Immunofluorescence for HA-tagged ETV3 expressed in HEK 293T cells showing nuclear localization regardless of ERK activity.
  • Fig. S5. Immunofluorescence for endogenous ETV3 in DLD1 cells showing primarily nuclear localization regardless of ERK activity.
  • Fig. S6. ETV3 binding sites occur near known transcriptional start sites.
  • Fig. S7. Phosphorylation by ERK2 does not affect the stability of ETV3.
  • Spectra. Manually curated spectra used to identify all AS-ERK2 targets.

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 20.2 MB

Other Supplementary Material for this manuscript includes the following:

  • Table S1 (Microsoft Excel format). AS-ERK2 substrates and signal-to-background ratios.
  • Table S2 (Microsoft Excel format). MEK-dependent phosphorylation in peptides isolated by phosphorylation motif peptide immunoprecipitation.
  • Table S3 (Microsoft Excel format). Overview statistics of ETV3 ChIP-Seq data.
  • Table S4 (Microsoft Excel format). ETV3 binding sites from ChIP-Seq.
  • Table S5 (Microsoft Excel format). Gene mappings for ETV3 ChIP-Seq binding sites.

[Download Tables S1 to S5 (Compressed)]

Technical Details

Format: Microsoft Excel. File is packaged as a compressed archive, in *.zip format; users should download the compressed file to their machine and decompress the file on their local hard drive, using the instructions below.

Size: 1.5 MB (compressed); 1.6 MB (decompressed)

Instructions for downloading and decompressing files:

  1. Create a temporary folder on your machine's hard drive.
  2. Save the compressed archive to the temporary folder you created, using the links above.
  3. Decompress the compressed file in the temporary folder using decompression software such as WinZip (Windows; or StuffIt Expander (Windows and Mac;

Citation: S. M. Carlson, C. R. Chouinard, A. Labadorf, C. J. Lam, K. Schmelzle, E. Fraenkel, F. M. White, Large-Scale Discovery of ERK2 Substrates Identifies ERK-Mediated Transcriptional Regulation by ETV3. Sci. Signal. 4, rs11 (2011).

© 2011 American Association for the Advancement of Science