Additional file 2 Figure S2.
p- Akt signal is abolished by phosphatase. The cells were starved as in Figure 2 and then stimulated by addition of insulin (1 mM) for 1 h. The cells were lysed in a buffer without EDTA and proteases inhibitors. Portions of lysates (200 μg of total protein) were incubated with calf intestinal alkaline phosphatase (0.05 U/mg protein) for 1 h at 37°C. The lysates were analyzed by Western blotting for Phospho Ser473-Akt. (PPT 114 kb)
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Gaben et al. BMC Cancer 2012 12:291 doi:10.1186/1471-2407-12-291