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. 2021 Jul 6;19(10):2069–2081. doi: 10.1111/pbi.13637

Figure 6.

Figure 6

ZmbHLH124 directly activates expression of ZmDREB2A. (a) Transactivation assay in tobacco leaf by transient expression. Left: firefly luciferase activity detection; middle: the relative expression levels of ZmbHLH124; right: PCR‐DNA amount of promoters to represent the equal plasmid DNA in agro‐infiltration were applied between parallel experiments, and NbACT was employed as the internal control in the DNA analysis. The data were based on three independent biological replicates. (b) Schematic diagram of the ZmDREB2A promoter. Red boxes indicate the putative E‐box motifs. (c) ChIP‐qPCR identified significant enrichment in the fragment containing the E‐box motif in the ZmDREB2A promoter. The fragments used in ChIP‐qPCR are indicated in (b). Fold enrichments were calculated relative to input. (d) ZmbHLH124T‐ORG directly bound to the promoter of ZmDREB2A in vitro. Lane 1–4: control; lane 5–10: protein and DNA interaction; lane 11–16: protein and mutated DNA interaction. Mutations of binding sites are labelled in (b). mu: mutation. Error bars, s.d., calculated from the results of at least three independent experiments. Statistical significance was determined by a two‐tailed t‐test: **P < 0.01; n.s.: not significant.