Fig. 3.

Cells lacking Jjj1, a strictly cytosolic protein, accumulate half-mer ribosomes at 23°C. (A) Ten OD₂₅₄ units of lysate from WT cells grown at 30°C were centrifuged through a 5–50% sucrose gradient and the OD₂₅₄ monitored (Upper). Fractions collected were analyzed by immunoblotting for the presence of Jjj1, Zuo1, and Rpl3 (Lower). (B) Ten OD₂₅₄ units of lysate from the indicated strains grown at 23°C were centrifuged through 7–47% sucrose gradients, and the OD₂₅₄ was monitored. (C) Log-phase cells of the LMB-sensitive strain AJY1539 were incubated in LMB (0.1 μg/ml) for 30 min and then fixed with formaldehyde and subjected to indirect immunofluorescence microscopy by using antibodies specific for Jjj1 or Nmd3. Localization was then monitored by using anti-rabbit rhodamine-conjugated antibodies. DAPI was used to visualize nuclei.