FIG 2.

Luc-K12t and Luc-nonstop fragments are efficiently released from ribosomes. (A) The Luc-K12t fragment is efficiently released from ribosomes. Lysates (L) were separated into a cytosolic supernatant (C) and ribosomal pellet (R) by centrifugation through a high-salt sucrose cushion. The immunoblot was decorated with antibodies directed against luciferase (Luc), Sse1 (cytosolic marker) and Rpl24 (ribosomal marker). (B) The Luc-nonstop fragment is efficiently released from ribosomes. The experiment was performed as described above for panel A. (C) Quantification of 3 independent experiments as shown in panel B. The intensities of the cytosolic (C) plus ribosomal (R) luciferase bands were set to a total of 100%; the ribosome-associated Luc-nonstop fragment is given as a percentage of the total. Error bars represent the standard errors of the means. The P value was calculated via Student's t test.