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. 2004 Feb;24(3):1033–1043. doi: 10.1128/MCB.24.3.1033-1043.2004

FIG. 7.

FIG. 7.

Phosphorylated AML1 is degraded in a time-dependent manner in COS-7 cells. COS-7 cells were cotransfected with pME18S (lanes 1 and 2), pME-FLAG-AML1 (lanes 3 to 6), or pME-FLAG-S249/266A (lanes 7 and 8), together with pCMVMK; starved in medium containing 0.1% FCS; and treated for 5 min with 10 (lanes 3 and 4) or 100 (lanes 1, 2, and 5 to 8) ng of EGF per ml plus 10% FCS in the presence of 50 mM sodium fluoride as a phosphatase inhibitor. The cells were harvested after 10 (lanes 1, 3, 5, and 7) or 30 (lanes 2, 4, 6, and 8) min of EGF stimulation, and 30 μg of total cell lysates was subjected to SDS-PAGE, followed by Western blotting with the anti-FLAG or antiactin antibody. +, present; −, absent; arrows, phosphorylated and unphosphorylated AML1.