Ubiquitin-dependent transport from the Golgi to the vacuole. A-B: Schematic representation of the constructs RFP-TMD20 (A) and RFP-TMD20-Ub (B). Tobacco mesophyll protoplasts and leaf epidermal cells expressing the plasmids indicated below. RFP-TMD20 (C) exhibits a punctate pattern, while RFP-TMD20-Ub (D) is efficiently transported into the vacuolar lumen and also accumulates in ring-like structures after wortmannin-treatment (+WM; inset in D). Vacuolar transport of RFP-TMD20-UbΔGG (E) and RFP-TMD20-Ub is indistinguishable. F-H: Confirmation of the localization from C-E in leaf epidermal cells. Scale bars = 5 μm.