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. 2010 Apr 7;285(24):18806–18816. doi: 10.1074/jbc.M109.098079

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Cool-1 is a tyrosine phosphosubstrate. A, NIH 3T3 cells were serum-starved for 15 h and then stimulated with EGF for the indicated time periods. Endogenous Cool-1 was immunoprecipitated with an anti-Cool-1 antibody and detected with either an anti-phosphotyrosine (PY) antibody (top panel) or an anti-Cool-1 antibody (bottom panel). B, viral-Src-transformed NIH 3T3 cells were serum-starved for 15 h and then stimulated with EGF for the indicated time periods. Endogenous Cool-1 was immunoprecipitated with anti-Cool-1 antibody and then detected with either an anti-PY antibody (top panel) or an anti-Cool-1 antibody (bottom panel). C, viral-Src-transformed NIH 3T3 cells were transfected with Myc-tagged Cool-1, serum-starved for 15 h, and then stimulated with EGF for the indicated time periods. The Myc-tagged Cool-1 was immunoprecipitated with anti-Myc antibody and then detected with an anti-PY antibody (top panel) or an anti-Cool-1 antibody (bottom panel).