FIGURE 4.

Recruitment of Cdc48 on stalled 60S particles by Rqc1 and Ltn1 is essential to prevent aberrant protein aggregation. Wild-type and rqc1Δ strains expressing the TAP-NonStop reporter were grown with or without Cdc48 depletion (13.5 h with doxycycline). Polysome extracts were separated on a sucrose gradient (10–50%), and TCA-precipitated fractions were migrated on polyacrylamide gels and analyzed by Western blotting using antibodies against the TAP-NonStop (PAP) and rabbit polyclonal against Nog1 (1/10,000) as a loading control and a marker of the 60S-sedimenting fractions. Aggregated species of the TAP-NonStop accumulating in the stacking gel are annotated. NS, NonStop.