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. 2016 Dec 5;173(1):155–166. doi: 10.1104/pp.16.01024

Figure 3.

Figure 3.

SUF4 regulates EC1.1 in yeast and in planta. A to C, Yeast one-hybrid analysis of interactions between SUF4 and pEC1.1. A, The EC1.1 promoter was divided into two bait fragments, and arrows indicate primers used for bait construction. B and C, Transformed yeast strains with the proximal fragment of the EC1.1 promoter were grown on either permissive –His–Leu medium (B) or selective –His–Leu with 5 mm 3-AT medium (C). Sections 1 and 4, pGADT7 without any insert (negative control); sections 2 and 3, pGAD-SUF4. D, GUS staining of homozygous pEC1.1(-457)::GUS plants. All egg cells show reporter activity. E, SUF4 is important for EC1.1 promoter activity in planta. suf4-1 mutants were crossed with homozygous pEC1.1(-457)::GUS plants. In the F1 carpels, only 25%, instead of the expected 50%, of egg cells were GUS positive; therefore, pEC1.1(-457)::GUS activation relies on SUF4. Bar = 20 µm. F, All five EC1 genes are down-regulated in suf4-1 mutant pistils, as indicated by real-time RT-PCR analyses. To normalize the expression level, we used UBIQUITIN10 or ACTIN8 (data not shown). The expression of each EC1 gene has been calibrated to 1 in wild-type pistils (wt). G, The normal EC1 gene expression is restored in suf4-1 suf4-1 pSUF4::SUF4-GUS pSUF4::SUF4-GUS pistils. The expression of each EC1 gene has been calibrated to 1 in wild-type pistils.