MDA-468TR-PTEN (with and without 100 ng/ml doxycycline) and MDA-468TR-vector (with dox) were serum starved overnight and the following day treated for.

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MDA-468TR-PTEN (with and without 100 ng/ml doxycycline) and MDA-468TR-vector (with dox) were serum starved overnight and the following day treated for 1 h with the indicated concentrations of ZD1839, and cells were harvested rapidly on ice after a 10 nm EGF... MDA-468TR-PTEN (with and without 100 ng/ml doxycycline) and MDA-468TR-vector (with dox) were serum starved overnight and the following day treated for 1 h with the indicated concentrations of ZD1839, and cells were harvested rapidly on ice after a 10 nm EGF stimulus. Lysates were separated by SDS-PAGE and immunoblotted using the indicated antibodies. EGFR, MAPK, and Akt activity were assayed at 5 min after the EGF stimulus, whereas activity of the downstream targets GSK-3 and 4EBP1 were assayed at 30 min after the EGF stimulus. Qing-Bai She et al. Clin Cancer Res 2003;9:4340-4346 ©2003 by American Association for Cancer Research