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Fig. 3. Localization of the PH-GFP fusion proteins in wild-type PGCs. (A,B,C) Images are taken from 63x time-lapse movie showing PGCs labeled with PH^CRAC-GFP at the indicated time points. A PGC in (A) shows stable localization of PH^CRAC-GFP to the membrane (Movie S3 in supplementary material). (B) No differential membrane localization can be observed either prior to the formation of a protrusion (indicated by an arrow) or at the site of the protrusion itself (indicated by an arrowhead; Movie S3 in supplementary material). (C) No polarized membrane localization of PH^CRAC-GFP can be seen in a PGC undergoing directed migration relative to somatic cells (Movie S4 in supplementary material). The arrow marks the direction of migration of the PGC whereas the white symbol marks a neighboring somatic cell serving as a reference point. (D) A PGC showing a uniform membrane localization of PH^Akt-GFP, whereas the PGC in (E) shows weak and spotty membrane expression of PH^GRP1-GFP. The PGC in (F) shows uniform distribution of PH^TAPP1-GFP. (G) Non-confocal images of PGCs labeled with PH^Akt-GFP. The arrow marks a forming protrusion showing no increased fluorescence. Brighter fluorescence is mostly apparent at regions of cell-cell contact (Movie S5 in supplementary material).

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