CDF2 interacts with DCL1.

<p>(<b>A</b>) Yeast two hybrid assays show the interactions between CDF2 and DCL1-RBD or HYL1. Co-transformed yeast colonies were spotted on the selective SC medium minus Trp and Leu, and then grown on SC medium minus His, Trp, and Leu supplemented with 5mM 3-amino-1, 2, 4-triazole (3-AT). (<b>B</b>) GST and MBP pull-down assays show the interaction between CDF2 and DCL1. (<b>C</b>) Co-IP assay shows the interaction between CDF2 and DCL1. The protein extracts from 22-day-old <i>Arabidopsis</i> plants coexpressing <i>pCDF2</i>::<i>CDF2-HA</i> and <i>pDCL1</i>::<i>DCL1-YFP</i> were incubated with anti-HA–conjugated agarose. The pellet was analyzed by immunoblotting with anti-HA and anti-GFP antibodies. (<b>D</b>) Bimolecular Fluorescence Complementation (BiFC) assays show that CDF2 or the C-terminal fragment of CDF2 (CDF2-C) interact with DCL1/HYL1 in D-bodies, while no interactions were observed between CDF2 and DCL1-9 or the N-terminal fragment of CDF2 (CDF2-N) and DCL1. Scale bar = 10μm. (<b>E</b>) Yeast two hybrid assays show that the C terminal fragment of CDF2 interacts with DCL1-RBD. C1, aa 189–457; C2, aa 271–457; C3, aa 360–457.