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. 2012 Jan 18;109(5):1790–1795. doi: 10.1073/pnas.1118282109

Fig. 2.

Fig. 2.

N 21-nt secondary siRNAs are phased and require RDR6 and DCL4 for biogenesis. (A) Plot of secondary siRNAs (blue line) as the number of tobacco 21-nt sRNA raw reads (y axis) along the N gene (U15605 coordinates, x axis). A map of N (Fig. 1A) and N and nta-miR6019 sequences in black and red, respectively, are shown below. Horizontal brackets below N sequence indicate 21-nt siRNA phasing. nta-siRNAI and nta-siRNAII registers and coordinates are indicated in blue. (B) N siRNA length [nucleotide (nt), x axis] and number (raw reads, y axis) in wild-type tobacco sRNA library. (C) 5′-terminal nucleotide (x axis) of N 21-nt siRNAs (in raw reads, y axis). (D) Relative levels of nta-TAS3 siRNA2142, nta-siRNAI, and nta-siRNAII in TG34 (wt) and TG34::nta-amiR:RDR6 tobacco lines (rdr6), measured by quantitative miR-ID analysis. Data normalized to miRNA390 levels. Quantitative analysis was repeated with two biological replicas and three technical replicas for each. (E) Relative level of nta-TAS3 siRNA2142, nta-siRNAI, and nta-siRNAII in TG34 tobacco (wt) compared with levels in TG34::nta-RNAi:DCL2,DCL4 (dcl24). Data was normalized to miRNA390 levels in each library.