(A–L) Tonoplast localised SNAREs and marker protein NET4A under high auxin conditions. pUBQ10::VAMP711-YFP (A and B), 35S::SYP21-YFP (C and D), SYP22::SYP22-GFP (in syp22) (E and F) and NET4A::NET4A-GFP (G and H) expressing seedlings treated with DMSO (A, C, E, G) or NAA (B, D, F, H) (500 nM; 20 hr). (I and J) Estradiol does not affect VAMP711-YFP abundance. pUBQ10::VAMP711-YFP expressing seedlings treated with DMSO (I) or estradiol (J) (10 µM estradiol; 20 hr). YUCCA6 expression under control of an estradiol inducible promoter in pUBQ10::VAMP711-YFP expressing seedlings after DMSO (K) or estradiol (L) (10 µM; 20 hr) treatment. Propidium iodide (green) for decorating the cell wall was used for confocal imaging of atrichoblast cells. (M) Western-blot (anti-GFP) representing VAMP711-YFP, SYP21-YFP, SYP22-GFP and NET4A-GFP protein abundance after NAA (500 nM; 20 hr) and control treatment as well as corresponding alpha-tubulin abundance for normalization. (N) Mean grey value of vacuolar localised SNAREs and marker protein NET4A after auxin treatments (500 nM; 20 hr) compared to DMSO treatments. (O) Mean grey value of VAMP711-YFP after YUCCA6 induction (10 µM; 20 hr). (P) Western-blot quantification (mean grey values). n = 3 biological replicates each consisting of a pool of 40–50 roots. Error bars represent s.e.m. For statistical analysis DMSO and NAA treatments were compared. For confocal analysis (N-O): n = 32 cells in eight individual seedlings. Student's t-test p-values: ***p < 0.001. Scale bar: 15 µm.