Skip to main content
. 2014 Mar 27;10(3):e1004243. doi: 10.1371/journal.pgen.1004243

Figure 1. Up-regulation of NAC089 by ER stress is directly controlled by bZIP28 and bZIP60.

Figure 1

(A–B) Up-regulation of NAC089 in the wild-type (wt) plants by ER-stress inducers tunicamycin (TM, 5 µg/ml) and dithiothreitol (DTT, 2 mM) (A) and in the wt, bZIP28 single mutant (zip28), bZIP60 single mutant (zip60) or double mutant (zip28zip60) by TM (5 µg/ml) treatment (B). The expression of NAC089 is normalized to the expression of the internal control actin. (C–D) Transactivation of NAC089 promoter in the dual-luciferase leaf protoplast assays. Activations of the NAC089 promoter by ER stress treatments (C) and by co-expression of either activated bZIP28 (bZIP28D) or activated bZIP60 (bZIP60S) (D). Relative reporter activity is the firefly luciferase activity normalized to the renilla luciferase activity. Bars depict SE (n = 3) in A–D. The empty vector (EV) was used as a negative control. ** P<0.01, * P<0.05. (E–F) EMSA experiments to detect the protein-DNA binding. Either the purified His-bZIP28D (E) or Trx-His-bZIP60T (F) was incubated with the biotin-labeled pNAC089 DNA. Lane 3, 50× un-labeled pNAC089; lane 4, 200× un-labeled pNAC089; lane 5, 200× un-labeled mutated form pNAC089M1. Arrows and arrow heads point to the positions of shifted bands and free probes, respectively.