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. Author manuscript; available in PMC: 2011 Feb 16.
Published in final edited form as: Dev Cell. 2010 Feb 16;18(2):226–236. doi: 10.1016/j.devcel.2009.11.015

Figure 2.

Figure 2

PHAKT-EGFP translocates to the leading edge when neutrophils come to and leave laser-induced wounds.

(A) Time-lapse ratiometric imaging (PHAKT-EGFP/mCherry) reveals PI(3,4,5)P3-PI(3,4)P2 localization at the leading edge during attraction to a laser wound in the tail fin (movie S2A). The white dots and arrows indicate the position of the wound and direction of migration respectively. (B) Reversal of PI(3,4,5)P3-PI(3,4)P2 (ratiometric imaging of PHAKT-EGFP/mCherry) when a neutrophil leaves the laser wound (movie S2B). Note loss of PI(3,4,5)P3-PI(3,4)P2 polarity at the wound, followed by reversal of polarity to the opposite pole away from the wound when the neutrophil leaves the wound (green line: tracking of a neutrophil, yellow thunder: position of the laser wound, white arrows: direction of migration, illustration with white line: morphology of a neutrophil). Data are representative of more than 5 time-lapse movies from a minimum of 3 separate experiments. The numerical values of ratiometric analysis are shown in the scales. Scale bars, 10 μm (A, B).