1. Prefoldins 3 and 5 Play an Essential Role in Arabidopsis Tolerance to Salt Stress 
Rodríguez-Milla Miguel A. , Salinas Julio  
Molecular Plant 
Volume 2, Issue 3, Pages (May 2009)
DOI: /mp/ssp016
Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

2. Figure 1
Complementation of Yeast gim Null Mutants with Arabidopsis PFD/GIM cDNAs.
Arabidopsis thaliana GIM1/PFD6, GIM2/PFD3, GIM3/PFD4, GIM5/PFD5, and GIM6/PFD1 were cloned into the low-copy-number plasmid pDR195 and expressed in yeast mutants gim1Δ, gim2Δ, gim3Δ, gim5Δ, and gim6Δ, respectively. BY4741 wild-type (WT) and gimΔ mutant cells transformed with the empty vector pDR195 were used as controls. Cells grown to mid-log stage in drop-out SD-URA medium were collected, and serial dilutions of the same number of cells spotted on YEPD plates (control) or YEPD plates supplemented with 1.2 M NaCl (1.2 M NaCl) and grown at 30°C.
Molecular Plant 2009 2, DOI: ( /mp/ssp016)
Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

3. Figure 2
Identification and Morphological Characterization of Arabidopsis pfd3 and pfd5 Mutants.
(A) Structure of Arabidopsis PFD3 and PFD5 genes, and T-DNA insertions in the pfd3 and pfd5 mutants. Exons are represented as gray boxes and introns as black boxes. The position and orientation of the T-DNA insertion in pfd3 (GABI-KAT line CR403489) and pfd5 (GABI-KAT line CT955299) mutants are indicated. ATG and TGA represent the start and stop codons, respectively.
(B) Identification of PFD3 and PFD5 cDNAs in pfd3 and pfd5 mutants by RT–PCR. RNA was extracted from leaves of wild-type (WT), pfd3, and pfd5 plants. The predicted products corresponding to PFD3 and PFD5 were obtained from wild-type plants but not from pfd3 or pfd5, respectively. PFD4 was used as a control.
(C) Morphological phenotypes of pfd3 and pfd5 mutants at different stages of development. Pictures show 2 (top) and 5 (bottom)-week-old wild-type (WT), pfd3, and pfd5 plants grown at 20°C and 16 h light.
(D) Cell shape in pavement cells from cotyledons of wild-type (WT) and pfd5 seedlings as shown by scanning electron microscopy. Scale bars  =  200 μm.
Molecular Plant 2009 2, DOI: ( /mp/ssp016)
Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

4. Figure 3 Microtubule Organization in pfd3 and pfd5 Mutants.
Organization of microtubules in root-elongating cells from 5-day-old wild-type (WT), pfd3, and pfd5 seedlings. DM1A anti-α-tubulin primary antibody and Alexa Fluor 488-coupled secondary antibody were used to visualize cortical microtubules. Fluorescence-labeled cortical microtubules were visualized with a confocal laser microscope. Scale bars  =  10 μm.
Molecular Plant 2009 2, DOI: ( /mp/ssp016)
Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

5. Figure 4 Tubulin and Actin Levels in pfd3 and pfd5 Mutants.
Levels of α- and β-tubulin and actin proteins in 3-week-old wild-type (WT), pfd3, and pfd5 plants. Soluble proteins were separated by SDS–PAGE and analyzed by Western blot using anti-α-tubulin, β-tubulin, and actin monoclonal antibodies. Coomassie-stained rubisco is shown as a control for equal loading.
Molecular Plant 2009 2, DOI: ( /mp/ssp016)
Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

6. Figure 5
Tolerance of pfd3 and pfd5 Mutants to Salt and Osmotic Stresses.
(A) Five-day-old wild-type (WT) and pfd3 and pfd5 seedlings grown in GM medium were transferred to vertical plates containing GM medium (control) or GM supplemented with 100 mM NaCl, 12 mM LiCl, or 300 mM mannitol, and allowed to develop for two (control seedlings) or three (treated seedlings) additional weeks.
(B) The tolerance of pfd3 and pfd5 mutants to NaCl was estimated by determining the fresh weight of seedlings after being transferred to a medium containing 100 mM NaCl as indicated in (A). The values are represented as the percentage of fresh weight of pfd3 and pfd5 seedlings with respect to the fresh weight of wild-type seedlings (WT) grown under control conditions or salt stress. Data are means of three independent experiments ± SD. For each experiment, 16 seedlings were pooled and weighted.
Molecular Plant 2009 2, DOI: ( /mp/ssp016)
Copyright © 2009 The Authors. All rights reserved. Terms and Conditions