DCP1 colocalizes with the SCAR–WAVE components SCAR2 and BRK1. Representative confocal micrographs showing colocalization between DCP1 and SCAR2 or DCP1 and BRK1 (in lines coexpressing DCP1pro:DCP1‐GFP and SCAR2pro:SCAR2‐mCherry or RPS5apro:HF‐mScarlet‐DCP1 and BRK1pro:BRK1‐YFP; arrowheads indicate colocalization at cell edges or vertices) in root epidermal cells (regions 2–3). Right: relative signal intensity profiles of DCP1 or SCAR2 at vertices. Colocalization at these regions was also calculated (PCC; N, biological replicates = 2, n = 4–10 edges/vertices in regions 2–3; a slight increase was observed in HS, 0.78 in NS vs. 0.87 in HS). Scale bars: 10 μm.
Representative confocal micrographs with acceptor photobleaching‐FRET efficiency between SCAR2‐mCherry (up) or BRK1‐mRuby (down) and DCP1‐GFP (epidermal cells, regions 1 and 3). Right: normalized FRET efficiency between SCAR2 or BRK1 and DCP1, respectively, among the different developmental root regions (N, biological replicates = 2, n = 16 cells). Scale bars: 3 μm.
Representative confocal micrographs showing PLA spots produced by α‐GFP/α‐RFP in DCP1‐GFP/SCAR2‐mCherry lines and DCP1‐GFP/BRK1‐mRuby lines, α‐FLAG/α‐GFP in HF‐mScarlet‐DCP1/SPI‐YPet (SPIRRIG [SPI] is a negative control as it localizes to PBs only during salt stress and was not found in the APEAL). In the SPI PLA, a high contrast inset is presented. Right: number of PLA spots per cell (N, biological replicates = 3, n (pooled data of 3 biological replicates) = 14–33 cells). In the merged images, the cell contours are shown (light green transparent). Scale bars: 5 μm.
Representative confocal micrographs showing DCP1 localization detected by α‐DCP1 in the wild type (WT) or the scar1 scar2 scar3 scar4 (scar1234) quadruple mutant or in live‐cell imaging of mScarlet‐DCP1 (RPS5apro:HF‐mScarlet‐DCP1) in WT or brk1 mutant (bottom right; epidermal cells, region 3 for α‐DCP1 and 2 for live‐cell imaging). The arrowheads denote the lack of robust DCP1 localization in scar1234 at the edge/vertex. Small panels (insets) at right show details corresponding to the regions delineated by dashed lines, where arrowhead denotes the edge signal of DCP1 in WT; scale bars, 1 μm. Bottom: percentage of cells with proper edge/vertex localization and quantification of PB numbers (DCP1‐foci; N, biological replicates = 3, n (pooled data of 3 biological replicates) = 18–35 cells). Scale bars, 5 μm.