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. 2005 Feb;25(3):907–920. doi: 10.1128/MCB.25.3.907-920.2005

FIG. 3.

FIG. 3.

RSK binds to NFATc4. (A) Endogenous NFATc4 were immunoprecipitated (IP) from untreated (−) or treated (+, Ion+PMA) mouse embryonic fibroblast cell extracts. Endogenous RSK in the immunoprecipitates was detected by immunoblot (IB) analysis. Activation and expression of RSK and NFATc4 in the cell extracts is also shown. (B) Extracts prepared from COS cells transfected with epitope-tagged NFATc4 and/or RSK2 were immunoprecipitated (IP) with NFATc4 or RSK2 antibodies. Associated RSK2 or NFATc4 was detected by immunoblot (IB) analysis. (C) Identification of RSK binding site on NFATc4. Various COOH-terminal truncated NFATc4 proteins (residues 1 to 902, 1 to 853, 1 to 581, 1 to 450, 1 to 365, 1 to 308, and 1 to 260) were coexpressed with epitope-tagged RSK2 in COS cells. NFATc4 proteins in the extracts and immunoprecipitated (IP) with RSK2 were detected by immunoblot (IB) analysis with M2 monoclonal antibody to the FLAG epitope. A schematic illustration of the NFATc4 protein and the RSK binding site is also shown.