Fig. 8: WAV3/WAVH function in polar PIN2 sorting in cytokinetic root meristem cells.

PIN2:Dendra signals in dividing cells of 5 DAG KN::PIN2:Dendra and wav triple KN::PIN2:Dendra. Arrowheads indicate reporter accumulation at the cell plate (a) and at apical and basal domains, respectively (b). Open arrowheads: limited signal accumulation at the wild type basal domain and wav triple apical domain. c PIN2-Dendra signals in dividing wild type (w.t.) KN::PIN2:Dendra (top) and wav triple KN::PIN2:Dendra (bottom) root meristem epidermis cells, after treatment with 50 μM MG132 for 2 h. Open arrowheads indicate intracellular signal accumulation, closed arrowheads indicate polar localization at the PM. d Quantification of PIN2-Dendra polarity in wild type (n = 24/14 roots) and wav triple (n = 22/19 roots) dividing root meristem cells at 5 DAG; n = 22–24. e PIN2:Dendra signals in KN::PIN2:Dendra and wav triple KN::PIN2:Dendra, after treatment with 10 μM BFA for 16 h. Arrowheads indicate apical localization of reporter signals. Open arrowheads: limited signal accumulation at the wild type basal domain and wav triple apical domain. f KN::PIN2:Dendra photoconverted before polarity re-establishment in wild type (w.t.; top) and wav triple (wtrp; bottom), to follow the fate of de novo synthesized PIN2 for 120 min. Arrowheads indicate polar localization of de novo synthesized PIN2 at apical and basal PM domains. Open arrowheads: limited signal accumulation at the wild type basal domain and wav triple apical domain. ‘de novo’ indicates protein accumulating after conversion (green); ‘conv.’ represents the converted protein fraction (magenta). Open arrowheads point towards minor signal accumulation at the basal domain in wild type and the apical domain in wav triple cytokinetic cells (b, e, f). Three independent experiments were performed for (a–c) and (e, f) with similar results. Circles represent single data points; boxes: first and third quartiles; center line: median; ‘x’: mean value. Two-tailed t-test was employed to determine statistical significance (d); ***: p < 0.001. Scale bars: a–c, e = 10 μm; f = 5 μm. Source data are provided as Source Data file.