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. 2013 Apr 25;9(4):e1003465. doi: 10.1371/journal.pgen.1003465

Figure 2. ADR1-L2 is required at the site undergoing cell death.

Figure 2

(A) Schematic of the chimera. adr1-L2 lsd1-2 expressing an estradiol inducible C-terminal HA-tagged ADR1-L2 were infiltrated in the indicated area with 20 µM estradiol, making that portion of the leaf ADR1-L2 lsd1-2. (B) Western blot to confirm expression of ADR1-L2 was limited to the estradiol-induced area. Estradiol + and − leaf areas were cored and protein was extracted from these cores. Protein extracts were run on SDS-Page gels and immunoblotted with anti-HA antibody. Coomassie stained blot confirms equal loading control (bottom). C, samples from un-infiltrated leaves; +, estradiol-infiltrated plant tissue; −, un-infiltrated tissue from the same leaf. In all samples, the entire leaf was treated with 300 µM BTH. (C) Trypan blue staining (top) of representative leaves (bottom) to show cell death in lsd1 control and tissue chimera plants. Leaves from four-week-old plants were treated as indicated in (A). Plants were sprayed with BTH 16 hours after estradiol treatment, and leaves were stained with trypan blue 5 days after BTH treatment.