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Fig. 3. PIKfyve serine318 phosphorylation is stimulated by constitutively active PKB and PtdIns 3-kinase mutants. Cells were transiently transfected with wild-type GFP-PIKfyve and vector alone, constitutively active PI3-kinase catalytic subunit (p110*) or constitutively active PKB (Myr-PKB). 48 hours later the cells were serum starved and then incubated in the absence or presence of insulin for 15 minutes, as indicated. Cell lysates were then subjected to western blotting with either the anti-pS318 or anti-PIKfyve antibodies as shown. The data are representative of three separate experiments.
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