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. 2019 Oct 10;9:14527. doi: 10.1038/s41598-019-50742-8

Figure 5.

Figure 5

Akt/PKB heterodimerisation with PDK1 and downstream activation correlates with the level of PDK1 homodimerisation in SKBR3 cells. (A) Representative intensity and FRET-FLIM images of SKBR3 cells co-transfected with mCherry-Akt/PKB and GFP-PDK1. (B) The efficiency of mCherry-Akt/PKB and GFP-PDK1 interaction determined by FRET-FLIM in SKBR3 cells increases upon growth-factor stimulation and is highest for R466A/K467A PDK1 mutant. (C) Phosphorylation of Akt/PKB at T308 correlates well with the efficiency of Akt/PKB and PDK1 heterodimerisation. (D) Phosphorylation of endogenous SGK1 at its T-loop by PDK1 is potentiated by the mutation R466A/K467A. (A) Scale bar: 20 µm. (B) N > 30. Box: 2xSEM; Whiskers: 80% population. Mann-Whitney test *p < 0.05. Three independent experiments. (CD) n = 3. Error bars: SEM.