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. 2016 Nov 28;6:37943. doi: 10.1038/srep37943

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Copyright © 2016, The Author(s)

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(a) The interaction of AhNAC2 and AhAREB1 was confirmed by yeast two-hybrid assays. Bait (AhAREB1) and prey (AhNAC2) were co-introduced by transformation into yeast strain AH109. The binding of the two TFs was confirmed by transferring the AH109 strain onto -Trp-Leu-His-Ade plus 5 mM 3-amino-1, 2, 4-triazole medium. WL: SD/-Trp/-Leu; WLHA: SD/-Trp/-Leu/-His/-Ade. (b) The physical interaction of AhNAC2 and AhAREB1 was confirmed by pull-down assays. Equal amounts of either GST or GST-AhAREB1 proteins were incubated with glutathione beads and then His-AhNAC2 protein was added for 4 h. After washing, samples were separated by 12% SDS-PAGE and identified by anti-AhNAC2 antibody. (c) AhNAC2 interacts with AhAREB1 in Arabidopsis protoplasts. AhAREB1 or AhNAC2 alone (AhNAC2-EYFP^C/EYFP^N, EYFP^C/AhNAC2-EYFP^N, AhAREB1-EYFP^C/EYFP^N and EYFP^C/AhAREB1-EYFP^N) were used as controls.