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. 2004 Apr 22;23(10):2116–2125. doi: 10.1038/sj.emboj.7600217

Figure 1.

Figure 1

Interaction of Fbw7 with c-Myc in vivo. (A) The Cdc4 phospho-degron (CPD) sequences of human cyclin E and c-Myc. The asterisk indicates L, I, or P; X indicates any residue other than K or R. (B) HEK293T cells were transfected with vectors for FLAG-c-Myc and either HA-Fbw1a, HA-Fbw2, HA-Fbw4, or HA-Fbw7, as indicated, and were then incubated with MG132 for 6 h. Cell lysates were subjected to immunoprecipitation (IP) with antibodies to FLAG, and the resulting precipitates as well as the original cell lysates (input) were subjected to immunoblot analysis (IB) with antibodies to HA or FLAG. (C) HEK293T cells were transfected with a vector for HA-Fbw7, subjected to serum deprivation, stimulated by re-exposure to serum for 2 h, and then incubated for 6 h in the additional presence of MG132. Cell lysates were then subjected to immunoprecipitation with antibodies to c-Myc or with control mouse IgG, and the resulting precipitates were subjected to immunoblot analysis with antibodies to HA or c-Myc.