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. 2021 Mar 26;12:1901. doi: 10.1038/s41467-021-22267-0

Fig. 1. Differential distribution of TGN-localized proteins.

Fig. 1

a–f Confocal images under conventional CLSM (a–c) or 3D images under SCLIM (d–f) of GFP-SYP61 and mRFP-SYP61 (a, d), GFP-SYP43 and mRFP-SYP61 (b, e), or GFP-SYP43 and VHAa1-mRFP (c, f) in the epidermal cells of the root elongation zone. g 3D colocalization analysis of the TGN markers: n = 64, 50, and 118 TGNs for GFP-SYP61 vs mRFP-SYP61, GFP-SYP43 vs mRFP-SYP61, and GFP-SYP43 vs VHAa1-mRFP, respectively, from five or more biological replicates. Two-sided Steel-Dwass test; P = 0.86 (Left: GFP-SYP61 × mRFP-SYP61 vs GFP-SYP43 × mRFP-SYP61), P = 3.0 × 10−14 (Top: GFP-SYP61 × mRFP-YP61 vs GFP-SYP43 × VHAa1-mRFP), and P = 2.2 × 10−7 (Right: GFP-SYP43 × mRFP-SYP61 vs GFP-SYP43 × VHAa1-mRFP); *P < 0.01, NS = nonsignificant. Boxes represent 25% and 75% quartiles, lines within the box represent the median, and whiskers represent the minimum and maximum values within 1.5× the interquartile range. h 3D images of GFP-SYP61, TagRFP-VAMP727, and iRFP-VAMP721 in the epidermal cell of the root elongation zone under SCLIM. i Multi-angle magnified 3D images of GFP-SYP61, TagRFP-VAMP727, and iRFP-VAMP721 of the boxed area in h. Upper panels: top view; middle and lower panels: side view (h, i). The experiments were repeated at least five times independently with similar results, and micrographs from representative experiments are presented. Scale bars = 5 μm (a–c); 1 μm (h, i). Grid width = 0.234 μm (d–f). Dashed lines indicate cell edges. V, vacuole area.