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. 2021 May 2;33(7):2320–2339. doi: 10.1093/plcell/koab120

Figure 7.

Figure 7

SlPIF4 directly regulates the expression of SlDYT1. A, EMSA showing the binding of the recombinant SlPIF4 protein to the promoter fragment of SlDYT1. DNA oligomers containing an E-box identified in the promoter region of SlDYT1 were labeled with digoxin and used as probes (P). mP1: mutant P1. SlPIF4937–1248, a truncated SlPIF4 protein containing the bHLH domain involved in DNA binding and protein oligomerization. B, Enrichment of the SlDYT1 promoter regions confirmed by ChIP-qPCR with the indicated primer sets (P1, P2, P3, and P4). Each error bar represents the mean ± sd, n = three biological replicates. Asterisk indicates a significant difference at P < 0.05 (Student’s t test). C, Y1H assay showing the binding of SlPIF4 protein to the promoter fragment of SlDYT1. The full-length coding sequence of SlPIF4 was fused to the GAL4 AD in the pGADT7 vector. 3*E: three tandem copies of E-box from the SlDYT1 promoter cloned into the pAbAi vector. Interaction was determined on SD/-Leu medium with AbA (-Leu+AbA300ng/ml). pSlDYT1, the SlDYT1 promoter.