Figure 2.

TOE1 mRNA Cleavage by miR172 in N. benthamiana Leaf Cells.

(A) Growth stage–dependent abundance of miR172 and its target gene transcripts. DCL1 and SE were also included for comparison in the assays.

(B) Abundances of TOE1 mRNA and miR172 in plant tissues. The 5S rRNA served as a loading control. CL, cauline leaves; F, flowers; RL, rosette leaves; FB, floral buds; R, roots; S, stems.

(C) Expression constructs of TOE1 genes and the MIR172a-2 gene. They were subcloned under the control of the CaMV 35S promoter.

(D) A truncated form (ΔTOE1) of the TOE1 gene used in the mRNA cleavage assays. It was used instead of a full-size one, since the miR172 cleavage site is located near the 3′ end. ORF, open reading frame; UTR, untranslated region.

(E) TOE1 mRNA cleavage by miR172 in N. benthamiana leaves. Cleavage products were examined by RNA gel blot hybridization using a TOE1-specific probe. Green fluorescent protein (GFP) transcription was also assayed to verify infiltration efficiencies. The arrow indicates the cleavage product derived from the 5′ side of the miR172 cleavage site.