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. 2021 Feb 8;118(8):e2010241118. doi: 10.1073/pnas.2010241118

Fig. 2.

Fig. 2.

LUX1 and LUX2 directly associate with the promoter of E1 to suppress its transcriptions. (A) Constructs of LUX1, LUX2, J, and E1 used for the transient expression assay in Arabidopsis protoplast. LUC, luciferase; REN, Renilla luciferase. (B) LUX1, LUX2, and J proteins suppress transcription from the E1 promoter in Arabidopsis protoplast. Values are shown as mean ± SD from three biological replicates. Different letters indicate significant differences by one-way ANOVA followed by Tukey’s post hoc test with SPSS statistics software. False-discovery rate (FDR)-adjusted P < 0.05. Two-way ANOVA revealed that the relative LUC activity is suppressed by J (P = 3.8 × 10−17), LUX (P = 1.2 × 10−19), and J × LUX (P = 3.9 × 10−9). (C) Schematic of the E1 gene and regions tested for enrichment in the ChIP assay and binding in the EMSA assay. (D) ChIP of E1 amplicons using W82, p35S:LUX1-FLAG, and p35S:LUX2-FLAG. Values are shown as mean ± SD from three biological replicates. Different letters indicate significant difference among the samples using the same primer by one-way ANOVA followed by Tukey’s post hoc test with SPSS statistics software. FDR-adjusted P < 0.05. Capital letters compare with each other, and lowercase letters compare with each other. (E and F) EMSA detected binding of GST-LUX1 (E) and GST-LUX2 (F) protein to the LBS of the E1 promoter.