Extended Data Fig. 4. Purification of a SpeFL-70S complex stalled in the presence of ornithine.

a, Overlaid absorbance profiles of sucrose gradients containing a translation mixture incubated without ornithine (black), in the presence of 10 mM L-ornithine (red) or in the presence of 10 mM L-ornithine followed by treatment with 100 μM puromycin (blue). This experiment was repeated three times independently with similar results. A schematic diagram depicting the expected ribosomal species in each fraction is shown on the right. b, Overlaid absorbance profiles of sucrose gradients loaded with polysomal fractions from a, with (blue) or without (black) RNase H treatment. This experiment was repeated three times independently with similar results. Expected ribosomal species for each fraction are shown on the right. c, Schematic representation of the purification strategy for SpeFL-70S. The collected fractions are indicated with gray boxes.