To discern for nonretrotranslocated and retrotranslocated ubiquitinated Hmg2-GFP, the substrate was immunoprecipitated from a 1× (24 μL) aliquot of the non-fractionated mixture (T) or from the supernatant (S) and pellet (P) of an identical volume (24 μL) of the same reaction mixture that was centrifuged, as described. Figure is taken from Neal, S., Mak, R., Bennett, E. J., & Hampton, R. (2017). A Cdc48 “retrochaperone” function is required for the solubility of retrotranslocated, integral membrane endoplasmic reticulum-associated degradation (ERAD-M) substrates. The Journal of Biological Chemistry 292, 3112–3128.