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. Author manuscript; available in PMC: 2010 Sep 25.
Published in final edited form as: Biochem Biophys Res Commun. 2009 Jul 16;387(3):521–524. doi: 10.1016/j.bbrc.2009.07.060

Figure 2. Carbachol induced Akt phosphorylation on Ser473 is EGFR-dependent but p70S6K phosphorylation on Ser389 is EGFR-independent.

Figure 2

A and B. T84 cells were incubated in the absence or presence of the EGFR inhibitor AG1478 (1 μM) for 1 h prior to stimulation of the cells with 100 μM carbachol (carb) or 50 ng/ml EGF for 10 min. The cultures were lysed with 2×SDS-PAGE sample buffer and analyzed by SDS-PAGE and immunoblotting with phospho Akt Ser473 and total Akt in panel A and phospho p70S6K Ser389 and total p70S6K to verify equal loading in panel B. Shown here are representative autoluminograms; similar results were obtained in 3 independent experiments. Autoluminograms were quantified by densitometric scanning. The results shown are the mean ± S.E.M. n=3 and are expressed as percentage of the maximum increase induced by treatment with carbachol, in cells preincubated in the absence (open bars) or the presence (closed bars) of AG1478 (panel C).