PotriNRAMP3.1 localizes to the TGN while PotriNRAMP3.2 localizes to the vacuolar membrane. (A) GFP translational fusions of PotriNRAMP3.1 and PotriNRAMP3.2 were imaged in vacuolar planes by spinning disk confocal microscopy in root epidermal cells (early elongation zone) of transgenic Arabidopsis T3 seedlings and poplar. NRAMP3.1-GFP labels granular cytoplasmic structures at the cell periphery, in both Arabidopsis and Poplar. In contrast, NRAMP3.2-GFP is present on the vacuolar membrane. Transmitted-light (differential interference contrast, DIC) and fluorescence (GFP) acquisitions are shown. (B) Colocalization of PotriNRAMP3.1-GFP with the TGN marker mRFP-SYP43 (top panel) and juxtaposition of PotriNRAMP3.1-GFP with the trans-Golgi apparatus marker mRFP-ST (bottom panel) in root epidermal cells (early elongation zone) of Arabidopsis F1 seedlings. Translational fusions were imaged in the cortical planes by spinning disk confocal. Note how PotriNRAMP3.1-GFP fluorescence either faces the center of the toroidal structure of the trans-Golgi or is present as Golgi-independent structures (white asterisks). On the merged images the overlap of GFP (green) and mRFP (magenta) channels appears white. Scale bar: 10 µm. Super resolution acquisitions (Super Res.) are 10 µm wide.