Figure 3.

Polysome analysis of TAS2-derived RNAs in wild-type and tasiRNA-defective mutants. A, Patterns of A₂₅₄ and distribution of ribosomal RNAs. An extract of floral tissues from wild-type (WT) plants was subjected to fractionation by 10% to 40% glycerol density gradient centrifugation. Absorbance was measured continuously. RNA was isolated from each fraction (1 mL), run on a 1.2% agarose gel, and stained with ethidium bromide. Fractions 1 and 12 represent the top and bottom fractions. 80S ribosomes were mainly fractionated in the fourth fraction. B, Polysome analyses of TAS2-derived RNAs in tasiRNA-defective mutants. Fractionation was performed as described in A. RNA blots were hybridized with the TAS2 probe. Total RNA (10 µg) from rdr6 mutant plants was used as a marker for the TAS2-related RNAs (the leftmost lane on each gel). Continuous absorbance measurement at 254 nm and RNA gels stained with ethidium bromide are shown in Supplemental Figure S3.