Localization of the ARF GTPase machinery at the PM. (A–C) Localization of the functional VAN3-GFP at the PM. (A) Heart-stage embryo. (B) Root cortex cells. (C) Lateral root primordium. (D–F) Confocal images of seedlings coexpressing (D) functional GNOM-GFP and (E) VAN3-mRFP, showing intracellular and PM localization and preferential PM labeling as well as minor dot-like structures, respectively. (F) Merged image of GNOM-GFP (green) and VAN3-mRFP (red). Inset shows nonoverlapping intracellular localization of GNOM and VAN3. (G–I) Confocal images of seedlings coexpressing GNOM-GFP and VAN3-mRFP in BFA-treated (40 μM, 1 h) root cells. (G) GNOM-GFP in BFA-treated cells revealing strong labeling of BFA compartments and the PM. (H) VAN3-mRFP in BFA-treated cells showing the PM signal. (I) Merged images of GNOM-GFP and VAN3-mRFP in BFA-treated root cells. Arrows and arrowheads mark the endosomal localization and the signal at the PM, respectively. [Scale bars: 10 μm (A), 12 μm (B), 20 μm (C), and 4 μm (D–I).]