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AMPylation of BiP is a reversible event. S2 cells were treated with 5 mm DTT for 1 h for ER stress induction. The cells were then washed and replated with fresh media to remove DTT and left untreated (A) or treated with 100 μg/ml cycloheximide (CHX; B). AMPylation of BiP, indicated by an arrowhead, was then monitored over time from the whole cell lysate using anti-AMP-Thr antibody.
