1. Volume 6, Issue 5, Pages 1605-1615 (September 2013)
The Lumen-Facing Domain Is Important for the Biological Function and Organelle-to- Organelle Movement of bZIP28 during ER Stress in Arabidopsis 
Le Sun, Sun-Jie Lu, Shuang-Shuang Zhang, Shun-Fan Zhou, Ling Sun, Jian-Xiang Liu 
Molecular Plant 
Volume 6, Issue 5, Pages (September 2013)
DOI: /mp/sst059
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

2. Figure 1 zip28zip60 Double Mutant Is Hypersensitive to TM.
(A) Phenotypes of zip28zip60 (dm) seedlings. Wild-type (WT) and dm were grown on ½ MS plates without or with various concentrations of tunicamycin (TM) for 10 d and then photographed.
(B, C) Effects of different concentrations of TM on root length (B) and leaf emergence rate (C) in WT and dm plants. Bars depict SE (n = 3).
Molecular Plant 2013 6, DOI: ( /mp/sst059)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

3. Figure 2
Either bZIP28 or bZIP60 Complements zip28zip60 (dm) Phenotype and Restores Up-Regulation of UPR Genes.
(A–C) Growth of transgenic plants complemented with different constructs. Coding sequence of bZIP28 (designated as LD1 in Figure 3) or bZIP60 was expressed in dm driven by its respective native promoter. Bars depict SE (n = 3).
(D) Validation of transgene expression under normal growth conditions. Primers were designed to amplify either the native gene or the transgene.
(E) Up-regulation of UPR marker genes in complemented transgenic plants.
Molecular Plant 2013 6, DOI: ( /mp/sst059)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

4. Figure 3
The Size of the bZIP28 Lumen-Facing Domain Is Critical for Its In Vivo Function.
(A) Diagram of constructs encoding full-length bZIP28 (LD1) and various deletion mutants (LD2–LD5). The transcriptional activation domain (TAD), DNA binding domain (DBD), transmembrane domain (TMD), three sequence conserved domains (CD1–CD3), and the S1P cutting site are shown above the LD1 illustration.
(B–D) Phenotypic analysis of transgenic plants in zip28zip60 (dm) background harboring various constructs (LD1–LD5), respectively. Bars depict SE (n = 3).
Molecular Plant 2013 6, DOI: ( /mp/sst059)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

5. Figure 4
The bZIP28 Lumen-Facing Domain Is Important for the Activation of UPR Downstream Genes.
(A) Validation of transgene expression in different transgenic plants under normal growth conditions. The primer was designed to amplify either the native gene or the transgene.
(B) Capability of different constructs to restore the up-regulation of UPR marker genes.
Molecular Plant 2013 6, DOI: ( /mp/sst059)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

6. Figure 5
The bZIP28 Lumen-Facing Domain Contains Subcellular Localization Signals.
(A) Diagram showing the GFP–myc fusions with different lengths of bZIP28 C-terminus. The bZIP28 N-terminus (aa 1–300) is replaced with GFP–myc in MLD1–MLD5. Abbreviations of TMD and CD1–CD3 are the same as in Figure 3.
(B–M) Confocal images of various chimerical proteins (MLD1–MLD5) in plants treated without (B–F) and with (G–M) DTT for 30min (G, I) or 4 h (H, J–M). Bar = 50 μm.
(N) Western blotting analysis of the processing of each fusion protein in response to DTT treatment (4 h). Asterisk denotes the position of the activated form. Coomassie blue staining of RbcS serves as a loading control.
Molecular Plant 2013 6, DOI: ( /mp/sst059)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

7. Figure 6
Hypothetical Model for the Organelle-to-Organelle Translocation of bZIP28.
(A) ER retention of bZIP28. Under normal growth conditions, the Golgi-localization signal (blue circle) is present at the cytoplasmic side of bZIP28. In the meantime, there is also a putative ER retention signal (blue triangle) on the bZIP28 C-terminus that dominates the Golgi-localization signal.
(B–D) Organelle-to-organelle movement of bZIP28. When misfolded proteins accumulate in the ER, the ER retention signal is abolished by an unknown mechanism so that the Golgi-localization signal becomes dominant (B). The bZIP28 is translocated from the ER to the Golgi where it is processed by S1P and S2P (C), leading to the nucleus accumulation and activation of downstream genes (D).
Molecular Plant 2013 6, DOI: ( /mp/sst059)
Copyright © 2013 The Authors. All rights reserved. Terms and Conditions