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. 2007 Apr 3;5(4):e95. doi: 10.1371/journal.pbio.0050095

Figure 3. Phosphoinositide-Dependent Change in PKB Conformation at the Plasma Membrane.

Figure 3

(A) NIH3T3 cells were transfected either with GFP-PKB or GFP-PKB-RFP. Upon PDGF treatment, GFP-PKB-RFP translocated to the plasma membrane and changed in conformation. The lifetimes are represented in pseudo-colour scale ranging from 2.20 to 2.63 ns. As indicated, the cells were pretreated for 20 min with LY294002 before PDGF stimulation. LY294002 prevented the translocation of GFP-PKB-RFP to the plasma membrane and its change in conformation but did not affect the FRET efficiency in the cytoplasm.

(B) The lifetime distributions are shown on the graphs: GFP-PKB control (blue), GFP-PKB-RFP (green), and GFP-PKB-RFP upon PDGF (orange) and treated with LY294002 before PDGF stimulation (dashed green). The separation of the lifetime pixels at the plasma membrane (red) and cytoplasm (light green) is shown for GFP-PKB-RFP upon PDGF stimulation.

(C) The statistical analysis presented as box and whiskers plots shows the extremely significant change in FRET efficiency (p = 0.0006) of GFP-PKB-RFP before and upon PDGF stimulation at the plasma membrane (PM) and the loss of FRET change when pretreated with LY294002.