The GAP domains of Leep2A and Leep2B are required for complex function. (A) TRITC-Dextran uptake in WT expressing GFP or DKO cells expressing GFP, GFP-Leep2A and Leep2B, GFP-Leep2^ΔGAP and Leep2B, or GFP-Leep2A and Leep2B^ΔGAP. (B) Quantification of TRITC-Dextran uptake, as shown in A. The scatter plot shows data points with means and SD; n represents the number of cells analyzed. Significance was determined by one-way ANOVA. (C) Quantification of TRITC-Dextran uptake by flow cytometry analysis. (D) Localization of GFP-Leep2^ΔGAP and RFP-Leep2B in DKO cells. (E) Localization of GFP-Leep2A and RFP-Leep2B^ΔGAP in DKO cells. (F) Co-IP of GFP, GFP-Leep2A, or GFP-Leep2A^ΔGAP with RFP-Leep2B or RFP-Leep2B^ΔGAP. Fluorescent fusion proteins were expressed in DKO cells. IP was performed with RFP-trap and samples were probed with GFP or RFP antibody. Scale bars, 5 μm. Source data are available for this figure: SourceData F6.