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Fig. 9. Effect of cAMP stimulation on PH-domain localisation. (A) Snapshots of aggregation competent cells just before and after cAMP stimulation (final concentration 5 µM). Only PtdIns(3,4,5)P3 binding PH domains show plasma membrane translocation. 20 to 40 cells were recorded for each cell line. (B) Kinetics of membrane translocation of the PtdIns(3,4,5)P3-specific probes. Data averaged from eight (CRAC) to ten cells (DAPP1/GRP1). (C) Membrane localisation of GRP1-PH and CRAC-PH during chemotaxis. Arrows indicate the direction of the cAMP source. (D) Changes in GRP1-PH-GFP localisation following repositioning of a cAMP-filled micropipette (position marked by arrows). See also supplementary material Fig. S6.





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