Fig. 4. centaurin-₂ interacts with the PI 3-kinase lipids products via the C-terminal PH domain in vivo. (A) Schematic of the mutant domains. (B) PC12 cells were transiently transfected with GFP, GFP-tagged centaurin-₂ (wild type; WT) or its mutants {R53C, the GAP mutant; R152C, the N-PH mutant; R276C, the C-PH mutant; R152C/R276C, the double PH domains mutant (DM)]. After 2 days, the cells were serum starved and live cells were imaged by confocal microscopy in the absence or presence of 0.1 µg/ml EGF. Images were captured 2 minutes after EGF addition. (C) The cells expressing GFP-tagged proteins were scored for the plasma membrane localisation. The number of cells (n) counted for each condition are shown in brackets in the graph. Data are from three experiments.