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. 2021 May 4;33(7):2431–2453. doi: 10.1093/plcell/koab122

Figure 2.

Figure 2

SYT3 localizes at ER–PM CS. A and B, SYT3-GFP shows a beads-and-string pattern in N. benthamiana leaves. Confocal images of lower epidermis cells transiently expressing SYT3-GFP and schematic representations of the cell planes that are showed in the images, either at the cortical region (A) or at the equatorial plane (B). Boxed regions are magnified in the insets (close up). See also Supplemental Figure S3, A and B. C, SYT3-mCherry colocalizes with ER–PM CS marker. Confocal images of N. benthamiana leaves transiently co-expressing SYT3-mCherry and the ER–PM CS marker MAPPER, showing the cortical region as in (A). Images of the individual channels as well as the merged are shown. See also Supplemental Figure S3, C and D. D, SYT3-GFP localizes at ER tubules and the edges of ER sheets. Confocal images of the lower epidermis cells of N. benthamiana leaves transiently co-expressing SYT3-GFP with the ER marker DGK2-mCherry. Images show the cortical region of the cells as in (A). Images of the individual channels as well as the merged images are shown. See also Supplemental Figure S3, E, F, and G. E and F, SYT3-GFP shows a beads-and-string pattern in Arabidopsis. Confocal images showing the localization of SYT3-GFP either at the cortical region of the cells (E) or at the equatorial plane (F) in epidermal cells from the cotyledon of 6-d-old Arabidopsis seedlings expressing 35S:SYT3-GFP grown in half-strength MS agar under long-day photoperiod and 23°C. Boxed regions are magnified in the insets (close up) G, SYT3-GFP colocalizes with endogenous SYT1 in Arabidopsis roots. Five-day-old Arabidopsis seedlings expressing 35S:SYT3-GFP grown in half-strength MS agar solidified medium under long-day photoperiod and 23°C were fixed with 4% PFA and permeabilized. The endogenous SYT1 protein was detected in the root tip with an anti-SYT1 primary antibody and a TRITC conjugated secondary antibody. Images of the individual channels as well as the merged image are shown. Scale bar 5 µm. See also Supplemental Figure S2G. A–F, Scale bars, 10 µm and scale bars for the close-up insets, 2 µm. A–G, Cortical plane images are a maximum Z-projection of several planes from the cell surface until a plane where cells are close but still not touching the neighbors. Equatorial plane images are single plane images Intensity plots along the white lines in close up views are shown for each co-localization pattern.