Polysome analyses support conclusions from Ribo-seq.

<p>Leaf lysates prepared from plants grown as for the ribosome profiling assays in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007555#pgen.1007555.g001" target="_blank">Fig 1</a> were fractionated by sedimentation through sucrose gradients. RNA extracted from each fraction was analyzed by RNA gel blot hybridization. The blots shown here come from material harvested at midday, either in the light or after 1 hour in the dark. A single blot for each tissue sample was probed sequentially to detect <i>rbcL</i>, <i>atpB/E</i>, and <i>psbA</i> RNAs. The methylene blue-stained blots below illustrate the distribution of cytosolic (25S and 18S) and chloroplast (16S and 23S*) rRNAs in the same gradients. The quantification shown at top is based on quantitative phosphorimaging. <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007555#pgen.1007555.s004" target="_blank">S3 Fig</a> shows the corresponding data for <i>ndhJ</i> and for material that was reilluminated for 15 minutes or harvested in the dark just prior to dawn.