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. 2014 Dec 30;26(12):4782–4801. doi: 10.1105/tpc.114.131607

Figure 6.

Figure 6.

Experimental Validation of NNC1 as a Target Gene of miR172c and Transcriptional Activity Analysis.

(A) and (B) Analysis of the cleavage of NNC1 by miR172c. The indicated constructs were transformed or cotransformed into N. benthamiana leaves, and the expression of NNC1 was imaged (A). Experiments were performed three times. Immunoblot analysis using antibody against GFP and relative GmNNC1-GFP accumulation in the different agroinfiltration assays are indicated in bar graphs below each panel (B). Experiments were performed three times. Different numbers indicate (as follows): 1, 35S:miR172c; 2, 35S:GmNNC1-GFP; 3, 35S:GmNNC1-GFP + 35S:miR172c; 4, 35S:GmNNC1m6-GFP; 5, 35S:GmNNC1m6-GFP + 35S:miR172c.

(C) Transcriptional activity of Gm-NNC1 was tested in N. benthamiana leaves using a GAL4/UAS-based system. 35S, the 35S promoter without the TATA box; 6×GAL4 UAS, six copies of the GAL4 binding site (UAS); G4DBD, the GAL4 DNA binding domain; G4DBD-GmNNC1, G4DBD fused with Gm-NNC1. Experiments were performed three times, and each experiment contained at least three replicates.