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. 2012 Jan 30;109(7):2461–2466. doi: 10.1073/pnas.1200169109

Fig. 2.

Fig. 2.

Secondary siRNAs triggered by 22-nt miRNA*s. (A) Small RNA blots to detect unmodified and modified miRNAs and miRNA*s expressed in transiently transformed N. benthamiana leaves. Secondary siRNAs produced from the TS:Luc reporter were also monitored. U6 was used as loading control. One asterisk (*) represents 21-nt miRNAs; two asterisks (**) represent 22-nt miRNAs. (B) Relative luciferase full-length mRNA levels, indicating effectiveness of trans silencing activity of secondary siRNAs. RNA levels were determined by quantitative RT-PCR using primers that did not amplify the TS:Luc fragment containing target sites for the miRNAs shown on top. Maximal silencing was defined using an artificial miRNA against luciferase (amiR:Luc); error bars are for SD. (C) Images of spatial distribution of luciferase intensity in N. benthamiana leaves. White, red, green, and blue indicate highest to lowest luciferase activity. Lower panel; leaves co-transformed with TS:LUC and miRNA constructs. (D) Quantification of luciferase activity in the same leaves.