Fig. 2.

The effect of UBR5A depletion on polysomes is more pronounced in rich media. UBHY-R cells were cultured in the indicated galactose-containing media (A and C) until reaching mid-log phase. Half of the culture was treated immediately with CHX and used to prepare a nondepleted extract. The other half was resuspended in the indicated media (B and D) with glucose and incubated at 30 °C for 5 (SD) or 6 (YP) h. After glucose incubation, extracts were prepared in the same manner. Approximately three OD units of each extract were loaded onto 10–60% (SD) or 10–50% (YP) sucrose gradients and centrifuged. Absorbance scans at 254 nm were obtained for each gradient. The ratio of polysomes to total ribosomes was calculated by numerical integration and is given for each profile (Upper Right). The profiles are representative of three independent analyses.