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. 2014 Oct 11;66(1):257–270. doi: 10.1093/jxb/eru415

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© The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 6. — Regulation of the PfCYCD2;1 expression. (A) Gene expression analysis of the putative PfCNR1 interacting partners and the putative Cyclin. Total RNA from the B2 flower buds was subjected to qRT-PCR. PfACTIN was used as an internal control. Expression of each gene was investigated in three independent biological samples and its expression in the wild-type P106 was set as 1. The mean and standard deviation are presented. (B) Characterization of the putative PfCYCD2;1 promoter. Three CArG-boxes (C1, C2, and C3) are highlighted in red circles beyond the ATG. The mutated promoters resulting from either a point mutation (C1pm) or deletion mutation (C1dm) are shown. The putative promoter sequences are presented in Supplementary Fig. S8. (C) Yeast one-hybrid assays between PfAG2 and the related DNA fragments. Pro, promoter; 3×, three tandem repeats of the CArG-box; C1, CArG-box1; C2, CArG-box2; C3, CArG-box3; dm, deleted mutation; pm, point mutation. The survival of yeast cells on SD/–Leumedium supplemented with aureobasidin A (AbA) suggested that the protein could bind to the corresponding DNA fragment. (D) LUC relative activity assay in P. floridana protoplasts. LUC activities were normalized using 35S:GUS as an internal control. The wild-type PfAG2 associated interactions are highlighted in pink and the interactions of PfAG2m that excluded the NLS are indicated in red. The mean and standard deviation from at least six independently repeated assays are presented. The star and the square, respectively, indicate a significant difference (P<0.05) compared with ProPfCYCD2;1:LUC or ProPfCYCD2;1:LUC with PfAG2. (E) The NLS in PfAG2 and its deleted version (PfAG2m). The 25 aa sequences of the NLS are given between the PfAG2 and PfAG2m structures. (F) Subcellular localization of PfAG2. The arrow indicates the nuclei. (G) Subcellular localization of PfAG2m. Bar, 10 μm (F, G).