Fig. 6: Wav triple-induced PIN2 polarity switches are mediated by BFA-sensitive cargo sorting.
From: WAVY GROWTH Arabidopsis E3 ubiquitin ligases affect apical PIN sorting decisions
a PIN2::PIN2:VEN reporter signal distribution in root meristem epidermis cells of 6 DAG eir1-4 (n = 47 cells/7 roots) and wav triple (n = 46 cells/8 roots) incubated in presence of 50 μM BFA for 90 min. Arrowheads indicate accumulation of reporter signals in BFA-induced intracellular compartments. b, c Subcellular PIN2 localization in wav triple root meristem epidermis cells in 6 DAG seedlings incubated on control plates (‘con.’; b) or in presence of 50 μM BFA (‘BFA’; c) for 16 h. Arrowheads indicate PIN2 polarity. d Orientation of root tips, expressed as degrees deviation from vertical of 6 DAG seedlings germinated on nutrient plates and then transferred on control nutrient plates (n = 81, wild type; n = 99, wav triple; n = 77, eir1-4 wav triple; n = 60, eir1-4) or on plates supplemented with 10 μM BFA (n = 85, wild type; n = 96, wav triple; n = 101, eir1-4 wav triple; n = 54, eir1-4) for 36 h. Subcellular localization of PIN2 in root meristem epidermis cells of 6 DAG wav triple (e; wtrp), gnomR5 (f) and wav triple gnomR5 (g). Arrowheads indicate PIN2 polarity. h Segregating wav triple and wav triple gnomR5 seedlings cultivated on vertically oriented nutrient plates. Open arrowheads indicate agravitropic wav triple primary roots, closed arrowhead indicates root of wav triple gnomR5. Arrow indicates the gravity vector. i Orientation of root tips, expressed as degrees deviation from vertical of 8 DAG wild type (n = 30), wav triple (n = 32), gnomR5 (n = 24) and wav triple gnomR5 (n = 26) seedlings cultivated on vertically oriented nutrient plates. Three independent experiments were performed for (b, c) and (e–g) with similar results. Circles represent single data points; boxes: first and third quartiles; center line: median; ‘x’: mean value. Two-tailed t-test (a, d) and Levene’s tests for the equality of variances (i) were employed to determine statistical significance; **: p < 0.01; ***: p < 0.001; a, b: p > 0.05. Scale bars: a–c = 10 μm; e–g = 20 μm; h = 10 mm. Source data are provided as Source Data file.
