Fig. 2

In vivo functionality of recombinant DCL4 in S. cerevisiae on an inducible hairpin transcript. A Positional sequence entropy plot of the hairpin transcript, 275-nt long constituted by gfp inverted repeats separated by the 79-nt RAD9 intron (with max entropy values). B Secondary structure and color map of base-pair probabilities of the hairpin transcript. C Relative expression level by qRT-PCR of the galactose-induced hairpin transcript in the absence and presence of copper-induced DCL4. Data are shown as the mean value SD from three independent assays using Student’s t-test (*P < 0.05; **P < 0.01). D Relative abundance of sRNAs from the SSC hairpin transcript, S. cerevisiae reference genome and transcripts. The relative abundance is also expressed in average part per million (ppm) referred to three replicates of sRNA sequencing. E Size distribution profile of 17-to-24 nt sRNAs from SSC hairpin transcript. F Size distribution profile of 20-to-24-nt sRNAs from TAS2 transcripts in A. thaliana Col-0 WT, and in mutant lines containing dcl2-4, dcl4-1 single mutants alleles and dcl4dcl2 double mutants alleles. G Distribution of 20-, 21- and 22-nt sRNAs (upper, middle and lower panels, respectively) along the hairpin transcript. Pink, violet and cyan lines refer to three independent replicates of short RNA sequencing. 5′-end positions of normalized sRNA reads were plotted using a square root scale. The red vertical line indicates the border of rad9 intronic loop