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. Author manuscript; available in PMC: 2013 Mar 30.
Published in final edited form as: Biochem Biophys Res Commun. 2012 Mar 3;420(1):161–165. doi: 10.1016/j.bbrc.2012.02.135

Figure 3.

Figure 3

Modulation of Akt signaling by DMI lacks selectivity. A. MEF cells were subjected to 5 minute stimulation with 10 μM NE alone, NE in combination with the α2AAR antagonist BRL44408, or NE following treatment with pertussis toxin (PTx) as described in the methods section. Whole cell lysates were analyzed by SDS-PAGE/Western blot for phospho-Akt and tubulin. Note that the lanes shown are from a single experiment but not all adjacent lanes. B. MEF cells were stimulated for 5 minutes with 10 μM NE alone (positive control) or 10 μM DMI alone and analyzed as in panel A. C. MEF cells were stimulated for 5 minutes with NE at varying concentrations, ranging from 10 nM (10−8 M) to 100 μM (10−4 M), alone or in combination with 10 μM DMI, as indicated, and analyzed as in panel A. D. Densitometric quantitation for panel A, with signaling activation calculated as a ratio of phospho-kinase:total protein. Data are mean ± SEM obtained over 3 independent experiments. *p<0.05 vs. NE alone. E. MEF cells were subjected to stimulation with 1% FBS for the indicated times, alone or in combination with 10 μM DMI, and analyzed as in panel A. All blots are representative of 3 independent experiments.