Figure S1.
Regulation of the membrane association of Leep1. (A) Time-lapse imaging of GFP-Leep1/pi3k1−2− cells during under-agarose chemotaxis. (B) Dd5P4− cells exhibited reduced TAPP1-GFP signal and dextran uptake. (C) Translocation of GFP-tagged TAPP1, cPH×3, and PHGRP1 in response to cAMP stimulation in the presence of LatA. (D) Sequential accumulation of GFP-Leep1 and RFP-cPH×3. GFP-Leep1 was selectively enriched at membrane ruffles and macropinocytic cups and quickly removed from internalized macropinosomes, whereas the signal of cPH×3 gradually increased until a peak was reached after the macropinosomes detached from the cell surface. (E) Sequence alignment of Leep1 with PH domains from CRAC or PkgE revealed a PH-like fold within the N terminus of Leep1. Cyan shading indicates the two positively charged residues (K28 and R39) mutated in Leep1AA. Scale bar = 10 µm.

Regulation of the membrane association of Leep1. (A) Time-lapse imaging of GFP-Leep1/pi3k1−2− cells during under-agarose chemotaxis. (B) Dd5P4− cells exhibited reduced TAPP1-GFP signal and dextran uptake. (C) Translocation of GFP-tagged TAPP1, cPH×3, and PHGRP1 in response to cAMP stimulation in the presence of LatA. (D) Sequential accumulation of GFP-Leep1 and RFP-cPH×3. GFP-Leep1 was selectively enriched at membrane ruffles and macropinocytic cups and quickly removed from internalized macropinosomes, whereas the signal of cPH×3 gradually increased until a peak was reached after the macropinosomes detached from the cell surface. (E) Sequence alignment of Leep1 with PH domains from CRAC or PkgE revealed a PH-like fold within the N terminus of Leep1. Cyan shading indicates the two positively charged residues (K28 and R39) mutated in Leep1AA. Scale bar = 10 µm.

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