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. 2013 Sep 26;8(9):e75773. doi: 10.1371/journal.pone.0075773

Figure 4. WUS binds the GRP23 promoter directly to activate its expression.

Figure 4

(A) qRT-PCR analysis of GRP23 expression in Ws, sef, Ler and wus-1. Data are means ± SD (n = 3). (B) qRT-PCR analysis of GRP23 expression in 14-day-old pga6 seedlings after inducing with 17-β-estradiol for 1, 3, 5 and 7 hours. Data are means ± SD (n = 3). (C) Transient expression assay in Arabidopsis protoplasts. The promoters of GRP23 and CLV3 were used to drive the luciferase (LUC) reporter gene. WUS:GFP fusion driven by the 35S promoter was used as effector. LUC activity was assayed after transformation. Data are means ± SD (n = 3). (D) ChIP assay of 7-day-old seedlings to show WUS-binding regions in GRP23 promoter. Regions i, -784 to -660; ii, -311 to -211; iii, -195 to -95. Data are means ± SD (n = 3). (E) EMSA of WUS binding the GRP23 promoter in vitro. The unlabeled double-strands probe (wt) and unlabeled mutant probe (mut) were used for competitive inhibition with 200X, 400X, or 800X molar excess.