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Sci. Signal., 25 October 2011
[DOI: 10.1126/scisignal.2002010]

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}mit.edu

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)


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  1. Create a temporary folder on your machine's hard drive.
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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


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