Identification of cargo proteins depending on the Gga1 Gga2 adaptor proteins. (a) Model highlighting the analysis of the GGA mutants within the CPY trafficking pathway. (b) Volcano plot identifying cargoes that are enriched or depleted at vacuoles of gga1Δgga2Δ cells. Analysis was performed as in Fig. 3 b. Vps10-dependent cargoes are labeled in green, depleted proteins in orange, and enriched plasma membrane proteins in blue. (c) Gga proteins affect the uptake of Mup1 from the plasma membrane. WT cells (upper panels) or gga1Δgga2Δ cells expressing Mup1-GFP were grown to the mid-log phase in SDC medium lacking methionine to induce Mup1 expression and retention at the plasma membrane. Cells were costained with FM4-64 to mark vacuoles and imaged 0 min, 30 min, and 60 min after the addition of methionine. Scale bar, 5 µm. (d) Localization of N-terminally GFP-tagged Sso1 and FM4-64–stained vacuoles was analyzed in WT cells (upper panel) and gga1Δgga2Δ cells (lower panel). Scale bar, 5 µm.