Figure 2
From: Fluorescence-based monitoring of ribosome assembly landscapes
Polysome analysis and fluorescence detection of sucrose fractions. Cells were grown in M9 medium at 37°C to OD600 = 0.4 and harvested. Lysates were subjected to sucrose gradient centrifugation. Centrifugates were analyzed by A254 detection and fractionated. Polysome profiles derived from: (A) MCrg*, (B) MCrg*ΔsQ, (C) MCrg*ΔlC. Sucrose gradient fractions of samples A-C were analyzed for mAzami- and mCherry specific fluorescence and normalized results are given in bar charts for: (D) MCrg* (E) MCrg*ΔsQ (F) MCrg*ΔlC. Superposition of A254 profiles and corresponding fluorescence bar charts: (G) MCrg*, (H) MCrg*ΔsQ, (J) MCrg*ΔlC. The inserts show fluorescence analysis of all available fractions from each sucrose gradient run. Red bars: normalized mCherry fluorescence; Green bars: normalized mAzami fluorescence. Fluorescence was normalized to the first polysome peak (“disome”) where subunits are supposed to be present in 1:1 ratio. Experiments were done in duplicates, representative profiles are shown.