Fig. 2.

The lipid signals and morphological factors underlying polarised accumulation of PH-PKB reporters. (A) An fMLP-containing micropipette was used to stimulate neutrophils co-expressing eGFP-PH-PKB and mCherry-CAAX, polar plots were created for each reporter (eGFP-PH – upper). (B) Validation of a PtdIns(3,4)P₂ reporter in neutrophils. Confocal images of neutrophils co-expressing mCherry-PH-TAPP-1 (upper) and eGFP-(R212L)PH-TAPP-1 (lower) stimulated with peroxy-vanadate for 0 or 170 s (representative of 4, see Fig. 3 for more frames from this movie and further controls). (C) The eGFP-PH-PKB construct reported changes in PtdIns(3,4,5)P₃ during neutrophil chemotaxis. An fMLP-containing micropipette (position out of figure directly down) was used to stimulate neutrophils co-expressing eGFP-PH-PKB (left) and mCherry-PH-TAPP-1 (right) (representative of 3).