Supplemental Data

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• Supplemental Data
• Supplemental Figure 1 - (A) WRKY proteins in parsley and Arabidopsis nuclear extracts. Nuclear extracts of parsley cell suspensions stimulated for 1 h with pep25 elicitor (lanes 1 and 3) or nonstimulated Arabidopsis cell suspensions (lanes 2 and 4) were analyzed by one-dimensional Western blotting using anti-all-WRKY serum in the presence of unspecific peptide (lanes 1 and 2) or immunogenic WRKY peptide (lanes 3 and 4). Molecular mass range is indicated on the right. (B) The anti-all-WRKY serum recognises different subgroups of Arabidopsis WRKY proteins. Different Arabidopsis WRKY proteins were expressed in E. coli either as amino-terminal GST-fusion or as amino-terminal 6x histidine-tagged proteins and total bacterial lysates analyzed by one-dimensional Western blotting using either the anti-all-WRKY serum (lanes 1-7) or commercially available anti-GST antibody (lanes 8-14). Lanes 1 and 8: pQE-His vector alone; lanes 2 and 9: His-AtWRKY6 (Group IIb); lanes 3 and 10: GST-AtWRKY11 (Group IId); lanes 4 and 11: GST-AtWRKY26 (Group I); lanes 5 and 12: GST-AtWRKY38 (Group III); lanes 6 and 13: GST-AtWRKY43 (Group IIc); lanes 7 and 14: pGEX vector alone. The anti-all-WRKY serum strongly detected multiple bands (due to the instability of the proteins) in all WRKY-expressing bacterial extracts but virtually no background signals in bacterial lysates harboring the vector control constructs. (C) Various modified forms of WRKY1 are detected by the anti-WRKY1 and anti-all-WRKY sera. Nuclear extracts of 3-h pep25 stimulated parsley cells were separated by two-dimensional PAGE and analyzed by Western blotting. Parallel gels were probed either with anti-WRKY1 (upper blot) or with anti-all-WRKY sera (lower blot). Arrows point to all WRKY1 signals that can be overlapped and are thus common to both blots. Molecular mass range is indicated on the right.