LC-MS/MS of co-immunoprecipitation of 3xFLAG-UFM1 from ribosomal pellet in control and upon anisomycin treatment

Clearance of arrested nascent polypeptides resulting from ribosomal stalling is essential for proteostasis. Stalled endoplasmic reticulum (ER)-bound ribosomes are marked by ubiquitin-fold modifier 1 (UFM1) on large ribosomal subunit protein RPL26, but the precise role of this process in ribosome-associated quality control (RQC) remains poorly understood. To identify proteins associated with UFMylated ribosomes, we isolated ribosomes from HCT116 cells expressing 3xFLAG-UFM1 upon stalling induction with anisomycin and in control conditions and preformed immunoprecipitation of 3xFLAG-UFM1.