Regulation of the localization and function of Leep1. (A and B) Time-lapse imaging of WT cells expressing RFP-Leep1 and PHcrac-GFP during under-agarose chemotaxis. RFP-Leep1 largely colocalized with PHcrac-GFP at pseudopods and during their retraction (indicated by the arrowheads) or splitting (indicated by the arrows). (C and D) Time-lapse imaging of GFP-Leep1/pirA− cells during under-agarose chemotaxis (C) or macropinocytosis (D). PirA was not necessary for recruiting Leep1 to pseudopods or macropinocytic cups. (E) GFP-ArpC4 localization during macropinocytosis. (F) Localization of RFP-Leep1 and GFP-ArpC4 at macropinocytic cups. (G) Localization of GFP-tagged chimeric proteins, which were generated by replacing the N terminus of Leep1 with PHcrac or PHPkgE. (H) Flow cytometry analysis of cells incubated with TRITC-dextran for 30 min. Scale bar = 10 µm. A.U., arbitrary units.
