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. 2015 Aug 3;5:12836. doi: 10.1038/srep12836

Figure 2. Accessibility of ribosome-stalled ubiquitin precursors to DUBs.

Figure 2

Mouse ubiquitin precursors were in vitro synthesized using stopless mRNAs (ΔSTOP). Synthesis was for (a) 30 min or (b) 45 min, in the absence or presence of HA-UbVME or HA-Ubal, as indicated. Aliquots of synthesis reactions performed in the absence of inhibitor were further incubated in the presence of either puromycin (Pur) or cycloheximide (CHX) for the indicated time periods. In (a) the peptidyl-tRNA species (UBA52-tRNA and UBA80-tRNA), unprocessed UBA52 and UBA80, as well as the corresponding processing products, L40, S27A and ubiquitin (Ub), are indicated. Note that S27A co-migrates with the abundant hemoglobin from the reticulocyte lysate hampering its detection. In (b) partially processed UBB and UBC species are indicated. Numbers to the left indicate the molecular weights of protein standards in kDa.