Fig. 8: Analyses of the interaction between the MdTMT1 promoter and MdMYB6 protein. | Horticulture Research

Fig. 8: Analyses of the interaction between the MdTMT1 promoter and MdMYB6 protein.

From: MdMYB6 regulates anthocyanin formation in apple both through direct inhibition of the biosynthesis pathway and through substrate removal

Fig. 8

a The 1520-bp sequence of the MdTMT1 promoter. b GUS histochemical staining of a callus harboring a plasmid containing the MdTMT1 promoter and GUS reporter gene (TMT1 pro: GUS) and a callus overexpressing MdMYB6 harboring the TMT1 pro:GUS vector (TMT1 pro: GUS + MYB6). c Analysis of GUS activity. d Yeast one-hybrid analysis of the interaction between the MdYB6 protein and the MdTMT1 promoter. e Predicted elements bound by MdMYB6 in the MdTMT1 promoter. f ChIP-PCR analyses of the interaction between the MdTMT1 promoter fragments and the MdMYB6 protein. MdMYB6-DNA complexes were then coimmunoprecipitated from the calli overexpressing MdMYB6-GFP via a GFP antibody. g EMSA analysis of the interaction between the MdTMT1 promoter and MdMYB6 protein. Lane 1 included labeled DNA probes with no added protein, while lane 2 included labeled DNA probes with no competitor or protein. Unlabeled DNA probes were added in increasing amounts (25×, 50×, and 100×) in lanes 3, 4, and 5. Unlabeled mutant DNA probes (lanes 7, 8, and 9) were added as competitors. Lane 6 included labeled mutant DNA probes and protein but without competitor.

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