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Figure 3


Fig. 3. The membrane recruitment of centaurin-{alpha}2 is mediated through PI 3-kinase activation. (A) PC12 cells transiently transfected with GFP-centaurin-{alpha}2 alone or with the PI 3-kinase regulatory subunit ({Delta}p85). After 2 days, the cells were serum starved and stimulated with 0.1 µg/ml EGF for 5 minutes. The cells expressing GFP-centaurin-{alpha}2 alone were preincubated with PI 3-kinase inhibitors (0.1 µM wortmannin or 50 µM LY294002) prior to EGF-stimulation. The cells were fixed and imaged by confocal microscopy. (B) PC12 cells were co-transfected with GFP-centaurin-{alpha}2 and Myc-tagged membrane-targeted PI 3-kinase (membPI3K) or its inactive form (kinase dead; membPI3K-KD). After 2 days, cells were serum starved, fixed, immunostained with an anti-Myc antibody (not shown) and imaged using a confocal microscope. (C) The EGF-stimulated cells expressing GFP-centaurin-{alpha}2 were scored for the plasma membrane localisation of centaurin-{alpha}2. The number of cells (n) counted for each condition are shown in brackets in the graph. Data are from three independent experiments.





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