Fig. 6: Verifying the regulatory pattern of TFs PyMYB114, PybHLH3, and PyWRKY26 in vivo.

a The dual-luciferase reporter assay verifying PybHLH3 and PyWRKY26 cotransactivation activity. b A schematic of the PyMYB114 promoter. The bar segments were analyzed by using the PLACE and PlantCare databases and are divided by solid lines (S1–S3). c Yeast one-hybrid assays between PybHLH3/PyWRKY26 and the PyMYB114 promoter. Positive colonies indicated strong specific interactions with the PyMYB114 promoter segments. The colony name indicates the bait segment, and prey vectors were used. d Model of the NLuc, CLuc and NLuc/CLuc constructs. e Firefly luciferase complementation assays in young tobacco leaves. Error bars are shown with six biological replicates. **P < 0.01. f Verification of the interaction of PyWRKY26 and PybHLH3 in vivo