ZFIN Image: Yoo et al., 2010, Fig. S3

Image

Figure Caption

Fig. S3 Imaging ofPI(3,4,5)P₃-PI(3,4)P₂ (related to Figure 3)

(A) PI(3)Kγ K799R expression impairs localization of PHAKT-EGFP at the leading edge, suggesting that PI(3)Kγ is primarily responsible for PI(3,4,5)P₃-PI(3,4)P₂ gradient formation.

(B) Expression of Δp85α, a deletion mutant of the adaptor subunit of class 1A PI(3)Ks, does not disturb localization of PHAKT-EGFP at the leading edge, suggesting that class 1A PI(3)Ks are not involved in PI(3,4,5)P₃-PI(3,4)P₂ gradient formation.

(C)PI(3,4,5)P₃-PI(3,4)P₂ pulse at the tail upon turning. Time-lapse ratiometric imaging of PHAKT-EGFP/mCherry at low magnification during locomotion of neutrophils in the mesenchymal tissues of the head (Movie S4B). PI(3,4,5)P₃-PI(3,4)P₂ pulse appears at the tail (magenta arrowhead) upon turning or changing directions in addition to the leading edge (green arrowhead). White arrows indicate direction of migration. Note that blue neutrophils have balanced expression of both PHAKT-EGFP and mCherry but that multi-colored neutrophils have high expression of PHAKT-EGFP with low or no expression of mCherry. Scale bars, 10 μm.

Acknowledgments

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Reprinted from Developmental Cell, 18(2), Yoo, S.K., Deng, Q., Cavnar, P.J., Wu, Y.I., Hahn, K.M., and Huttenlocher, A., Differential Regulation of Protrusion and Polarity by PI(3)K during Neutrophil Motility in Live Zebrafish, 226-236, Copyright (2010) with permission from Elsevier. Full text @ Dev. Cell