Fig. 3

OsSND2 directly regulate OsMYB61 expression. a Diagram of the effector construct and the reporter constructs used in b. b Yeast one-hybrid assay showing the activity of LacZ reporters driven by OsMYB61 and OsCESA11 promoters and activated by activation domain (AD) fusion effectors. The empty pB42AD and pLacZi were used as negative control. c Diagram of the effector construct and the reporter constructs used in d. d OsSND2 activates transcription of the OsMYB61 gene promoter. Relative luciferase activity was monitored in rice protoplasts cotransfected with different effector and reporter constructs. Mock, cotransfected with reporter construct and an empty effector construct; control, cotransfected with effector construct and an empty reporter construct (set to 1). Error bars, SE of three biological replicates. Student’s t-tests were used to generate the P values. The asterisks (**) indicate p < 0.01. e ChIP assays. The diagram depicts the regions used for ChIP-qPCR analysis of extracts from the second internodes of over-expression plants carrying the p35S::Myc-OsSND2 construct. ChIP-qPCR results were quantified by normalization of the Myc immunoprecipitation signal by the corresponding input signal. Error bars, SE of three biological replicates. f EMSA analysis. Competition for OsSND2 binding was performed with unlabelled P9 fragments containing SNBE1 site at 50×, 100× and 200× the amount of labeled probe