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. 2021 Jan 28;22(3):1297. doi: 10.3390/ijms22031297

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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SaARF4 directly downregulated SaACO4. Quantitative RT-PCR was conducted with three biological replicates and technical replicates to identify the effect of SaARF4 overexpression on the expression of SaACO4 (A). Vectors in the dual luciferase assay system were performed (B). The results of the Y1H indicated physical binding between SaARF4 and the SaACO4 promoter (C). HSa, pHIS2 + pGADT7-rec-SaARF4; HA, pHIS-AuxRE + pGADT7-rec; HASa, pHIS2-AuxRE + pGADT7-rec-SaARF4. The intensity of the fluorescence indicates the promoter activities of SaACO4 (D) in three biological replicates. Quantitative RT-PCR with Ren as the reference gene was conducted to further verify the results of the dual luciferase assay system (E,F). RML, relative mRNA level. * p < 0.05; ** p < 0.01.