Localization dynamics of the Leep2 complex. (A) Time-lapse imaging of WT cells co-expressing GFP-Leep2A, Leep2B, and PHcrac-RFP. Arrowheads indicate an emerging macropinosome. (B) Quantification of the changes in fluorescent intensity of PHcrac-RFP and GFP-Leep2A on newly formed macropinosomes, as shown in A. The frame at which the macropinocytic cup closed was set as time 0, and the fluorescence intensity at other time points was normalized to that at time 0. (C) Time-lapse imaging of WT cells co-expressing GFP-Leep2A, Leep2B, and TAPP1-RFP. Arrowheads indicate an emerging macropinosome. (D) Quantification of the changes in fluorescent intensity of TAPP1-RFP and GFP-Leep2A on newly formed macropinosomes, as shown in C. The frame at which the macropinocytic cup closed was set as time 0, and the fluorescence intensity at other time points was normalized to that at time 0. (E–G) Localization of GFP-Leep1 in WT (E), pi3k1⁻2⁻ (F), and pten⁻ (G) cells. (H–J) Localization of RFP-Leep2A and GFP-Leep2B in WT (H), pi3k1⁻2⁻ (I), and pten⁻ (J) cells. Scale bars, 5 μm.