Fig. 4: Interactions between OsWRKY53 and OsHKT1;5 in salt tolerance.
From: Genome-wide association studies identify OsWRKY53 as a key regulator of salt tolerance in rice
A Yeast one-hybrid assays to study interaction between transcription factor OsWRKY53 and target gene OsHKT1;5. B DNA binding activity of OsWRKY53 protein on OsHKT1;5 promoter fragment tested by EMSA assays. Brackets showed protein-DNA complex or free probe, respectively. EMSA assays were repeated three times. MBP-OsWRKY53 fusion protein was expressed in E. coli. The probes were labeled with biotin. C OsWRKY53 binding to W-box element of OsHKT1;5 promoter in surface plasmon resonance (SPR) experiments by using Biacore T200 instrument. D Different binding sites of transcription factor in the promoter of OsHKT1;5 and schematic representation of constructs used in transcriptional activity assay. E OsWRKY53 trans-repressed OsHKT1;5. Dual-luciferase reporter assay was used to measure luciferase activity. F Truncated promoters harboring different number of W-box were generated to drive firefly luciferase gene. Renilla luciferase gene was used as an internal control. Data were normalized to the internal control 35 S::REN. The values were significantly different from that of the control. Data were presented as means ± SD. P values were calculated with two-sided Student’s t-test, n = 3. Source data were provided as a Source data file.
