SE‐FRET efficiency between DCP1‐GFP or its phosphovariants with SCAR2‐mCherry (among the three different root regions; mainly epidermal cells). Scale bar: 50 μm. The arrowhead denotes high FRET efficiency at edges/vertices of region 3. Right: signal quantification of SE‐FRET efficiency between the indicated combinations at the epidermis of 3 regions (N, biological replicates = 6, n (pooled data of 3 biological replicates) = 10).
Actin nucleation site at an edge/vertex, as indicated by DCP1‐GFP and LifeAct‐mCherry localization. Right: correlation between DCP1 intensity, ACTIN, SCAR2, BRK1 and ARPC5 intensities (simple regression model). The R
2 values are shown, along with representative micrographs for DCP1/SCAR2 (N, biological replicates = 3, n = 6 for each point).
Representative confocal micrographs showing SCAR2‐mCherry localization in WT and dcp1‐3 mutant, respectively (root region 2, epidermal cells) and quantification of edge/vertex with SCAR2 a confined signal (N, biological replicates = 3, n = 5–8 cells).
Representative confocal micrographs showing the colocalization between DCP1‐GFP or phosphovariants and SCAR2‐mCherry in root meristematic cells (root region 2, epidermal cells). Scale bars: 10 μm. The insets show details of colocalization; the white arrowhead denotes the expansion of the SCAR2/DCP1 domain, while the yellow arrowheads the restricted edge/vertex SCAR2/DCP1 domains. Scale bars: 3 μm. The graph indicates the relative signal intensity for the indicated combinations (as Pearson's correlation coefficient; N, biological replicates = 3, n = 5 at edges/vertices: spread edges were not considered in calculations).