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Fig. 6 | BMC Biology

Fig. 6

From: Dynamic enhancer transcription associates with reprogramming of immune genes during pattern triggered immunity in Arabidopsis

Fig. 6

Validation of the expression dynamic and activation of predicted PTI upregulated enhancers. a Genome browser view of candidate transcribed enhancers. Pale blue bars indicate eRNA transcription regions. b eRNA induction level of candidate transcribed enhancers upon flg22 treatment was verified by RT-qPCR. Ten-day-old seedlings were treated with 100 nM flg22 for 1 and 3 h. The data are shown as mean ± SD from three independent repeats. Different letters indicate significant differences by the one-way ANOVA test (P < 0.05). c The activation in transcriptional regulation of candidate transcribed enhancers in transient expression assays. Arabidopsis protoplasts were infiltrated with constructs of pGL3B_Enhancer-mini35s::LUC. The y-axis represents the fold enrichment of luciferase signals of each construct compared to the control construct containing the mini 35S promoter. d The flg22 treatment increased activity of candidate enhancers. N. benthamiana leaf transiently transformed using Agrobacteria bacterial containing different constructs. Color scale represents the luminescent signal intensity measured by cps (counts per second). e WRKY33 is required for the activity of candidate PTI enhancers. Transient expression assays showing the function in promoting reporter gene expression of candidate enhancers in WT and wrky33 mutant. f The activity of candidate enhancer rely one W-box binding motif. Relative function of wild-type form enhancer and cognate w-box mutants were monitored in Arabidopsis protoplasts. Data are presented in c, e, f as the mean with standard error form six independent biological replicates. The P-values were based on a one-tailed Student’s t test (*P < 0.05, **P < 0.01; ns, no significance)

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