Detection of ribosome ubiquitination under normal and stress conditions. (A) Detection of ribosome ubiquitination. Total lysates (input) and FLAG-IPs obtained from WT and rpl36-FLAG embryos at 24 hpf were subjected to immunoblotting analysis with an anti-Ubiquitin antibody (left) and protein staining (right). (B) Analysis of ribosome ubiquitination under harringtonine (HTN) treatment. FLAG-IPs were subjected to immunoblotting analysis with an anti-Ubiquitin antibody (left) and protein staining (right). HTN concentrations are indicated above. (C) Analysis of ribosome ubiquitination under cycloheximide (CHX) treatment. FLAG-IPs were subjected to immunoblotting analysis with an anti-Ubiquitin antibody (left) and protein staining (right). CHX concentrations are indicated above. (D,E) Analysis of ribosome ubiquitination in the presence of mutant release factor eRF1-AAQ. Total lysates were subjected to immunoblotting analysis of eRF1-HA and Tubulin (D). FLAG-IPs were subjected to immunoblotting analysis with an anti-Ubiquitin antibody (E, left) and protein staining (E, right).
