Rab-A1c GTPase Defines a Population of the Trans-Golgi Network that Is Sensitive to Endosidin1 during Cytokinesis in Arabidopsis  Xingyun Qi, Huanquan Zheng  Molecular Plant  Volume 6, Issue 3, Pages 847-859 (May 2013) DOI: 10.1093/mp/sss116 Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 1 Rab-A Proteins Are Co-Localized to a Population of TGN. AGFP–RAB-A1c is co-localized with YFP–RAB-A4b. BGFP–RAB-A1c and YFP–RAB-A1b co-localize at a population of punctae. CGFP–RAB-A1c is co-localized with VTI12–YFP. White arrows indicate co-localized punctae. Pearson and Spearman correlation coefficients (PSCs) between GFP–RAB-A1c and each marker are indicated in the scatter plots on the right. Bars = 5 μm for all. Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 2 GFP–RAB-A1c Defined Punctae Are Distinct from Golgi and the Pre-Vacuolar Compartment but Partially Co-Localized with VHA-a1 Marked-TGN. AGFP–RAB-A1c punctae are associated but not completely co-localized with ST–YFP marked Golgi. BGFP–RAB-A1c and YFP–RAB-F2a label distinct punctae. CGFP–RAB-A1c only partially co-localize to VHA-a1-mRFP. Yellow arrow indicates GFP–RAB-A1c only punctae; red arrow indicates YFP–RAB-F2a only punctae. Pearson and Spearman correlation coefficients between GFP–RAB-A1c and each marker are shown in the scatter plots on the right. Bars = 5 μm for all. Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 3 Effects of ES1 and Wortmannin on the Subcellular Distribution of GFP–RAB-A1c. APunctate structures labeled by GFP–RAB-A1c in a wild-type cell. BGFP–RAB-A1c is relocated to a disc-like flat structure resembling the growing cell plate in mitotic cells in wild-type. CAggregation of GFP–RAB-A1c punctae in a cell treated with ES1. DAggregation of GFP–RAB-A1c punctae close to a cell-plate-like structure after ES1 treatment. The red arrow indicates the abnormal cell plate. EPunctae distribution of GFP–RAB-A1c is not affected by wortmannin. FGFP–RAB-A1c is still relocated to cell-plate-like structure when treated with wortmannin. Bars = 5 μm for all. Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 4 Promoter Activity of pRAB-A1a, pRAB-A1b, pRAB-A1c, and pRAB-A1i Reported by GUS Expression.Images of GUS staining of seedlings (A–D), roots (E–L), flowers (M–P), and pollen grains (Q–T) of transgenic plants expressing pRAB-A1a::GUS (A, E, F,M, Q), pRAB-A1b::GUS (B, G, H, N, R), pRAB-A1c::GUS (C, I, J, O, S), and pRAB-A1i::GUS (D, K, L, P, T). Note pRAB-A1b::GUS and pRAB-A1c::GUS have strong GUS activity in shoot meristems (B and C, red arrows), root tips (G and I, brackets), and at initiation of lateral roots (H and J, red arrows). pRAB-A1a::GUS also has GUS activity in root tips (E) and the initiation of lateral roots (F, red arrow). Bars = 1 mm for (A–D, M–P); 0.5 mm for (E–L); 0.2 mm for (Q–T). Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 5 Root Growth of rab-a1a/b/c Triple Mutant Is Slightly Retarded. (A) Stereo-microscopic image of 7-day-old seedlings of rab-a1a/b/c (left) and wild-type (right) (abc indicates rab-a1a/b/c triple mutant). Bar = 1 cm. (B) Quantitative analysis of 7-day-old seedlings of rab-a1a/b/c (n = 29) and wild-type (n = 32) (abc indicates rab-a1a/b/c triple mutant). Error bar represents standard error. Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 6 Different Effects of ES1 and Wortmannin on the Root Development of rab-a1a/b/c and Wild-Type Plants. (A–H) Seven-day-old seedlings of rab-a1a/b/c and wild-type grown on AT plates with 0 (A), 0.0835 μM (B), 0.167 μM (C), 0.33 μM (D) of ES1, and AT plates with 0 (E), 0.05 μM (F), 0.1 μM (G), 0.5 μM (H) of wortmannin. (I, J) Quantitative analyses of the reduction in root length in ES1-treated seedlings (I, n = 28–32) and wortmannin-treated seedlings (J, n = 28–32). The x-axis indicates concentration of drugs (μM), and the y-axis indicates the percentage of root length reduction relative to the root length of seedlings grown on AT plate with no drugs. Bars = 1.5 cm for all. Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 7 Cytokinesis in rab-a1a/b/c Triple Mutant Is Hypersensitive to ES1. (A–G) Calcofluor and propidium iodide stained cells in root tips of wild-type (A, D) and rab-a1a/b/c (B, C, E, F, G) growing on AT plates containing either DMSO (A–C) or 0.0835 μM ES1 (D–G). Note in rab-a1a/b/c, bi-nucleated cells were frequently observed when treated with ES1 (E–G, arrows), whereas, under the DMSO condition, bi-nucleated cells occur only occasionally (C, arrow). HQuantitative analyses of the number of bi-nuclei cells per root tip (n = 12) for each condition of both wild-type and rab-a1a/b/c. Bars = 5 μm for all. Error bar represents standard error. Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 8 Cytokinesis in Plants Expressing Dominant Inhibitory RAB-A1c Mutants Is Defective. (A–F) Seven-day-old seedlings of wild-type (A, D), transgenic plants expressing RAB-A1c(S27N) (B, E) and RAB-A1c(Q72L) (C, F) growing on AT plates containing either DMSO (A–C) or 20 μM dex (D–F). (G–I) Calcofluor and propidium iodide staining of root tip cells of wild-type (G), plants expressing RAB-A1c(S27N) (H) and RAB-A1C(Q72L) (I). Note bi-nucleated cells were frequently observed when the expression of dominant inhibitory RAB-A1c mutants is induced by dex (H and I, arrows). Bars = 1.5 cm for all. JQuantitative analysis of the root length (n = 28–32) of seedlings for each condition. Error bar represents standard error. Molecular Plant 2013 6, 847-859DOI: (10.1093/mp/sss116) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions