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Figure 5


Fig. 5. centaurin-{alpha}2 shows sustained kinetics of membrane recruitment after stimulation with EGF. (A) PC12 cells transiently transfected with either GFP-centaurin-{alpha}2, GFP-cytohesin 3, GFP-TAPP1 or GFP-PLC{delta}1 PH expression plasmid for 2 days were serum starved and live cells were imaged by confocal microscopy in the presence of 0.1 µg/ml EGF. Time points (top) indicate minutes of EGF stimulation. (B) PC12 cells were transiently transfected with expression plasmids encoding GFP-cytohesin 3, GFP-TAPP1 or GFP-centaurin-{alpha}2 with HA alone (vector), HA-PIPP or FLAG-4-phosphatase. After 2 days, cells were serum starved and stimulated with 0.1 µg/ml EGF for 2 minutes, fixed, immunostained with an anti-HA or an anti-FLAG antibody (not shown) and imaged using confocal microscopy. (C) The transfected cells, as shown in B, were scored for GFP-tagged protein as an indicator of plasma membrane localisation. A minimum of 200 transfected cells from three independent experiments were scored for each condition.





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