Fig. 3.

Lmo0762 is involved in recycling of antibiotic-stalled 70S ribosomes. Wild-type (blue) and Δlmo0762 (purple) bacteria were grown in BHI medium until exponential phase and erythromycin (0.18 µg/mL) was added or not for 1 h. Chloramphenicol was then added to the culture (2-min exposure at 5 mM) to stabilize the polysomes, and bacteria were pelleted and flash frozen. The cellular content was extracted, and equal amount of lysate (A₂₆₀) was loaded on a 5–50% sucrose gradient. After ultracentrifugation, the samples were collected from top (0 mm) to bottom (80 mm) of the tubes and the absorbance at 260 nm was monitored using a UV lamp. The baseline was corrected and the results were normalized based on the area under the curve.