Unanchored tri‐NEDD8 inhibits PARP‐1 to protect from oxidative stress‐induced cell death WT and NEDP1 KO HEK 293 cells were transfected with FLAG‐NSUN2.

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Unanchored tri‐NEDD8 inhibits PARP‐1 to protect from oxidative stress‐induced cell death WT and NEDP1 KO HEK 293 cells were transfected with FLAG‐NSUN2 and 48 h later harvested for anti‐FLAG immunoprecipitation. Following stringent washes, pulldowns were treated with recombinant GST‐NEDP1, the deubiquitinase his6‐USP1/UAF1, or left untreated. After incubation at 30°C for 30 min, reactions were stopped by the addition of LDS sample buffer, and then, samples were processed for Western blot analysis with the indicated antibodies. There is no shift in the anti‐FLAG immunoreactive band, which suggests NSUN2 is neither modified by NEDD8 or ubiquitin. IF THIS IMAGE HAS BEEN PROVIDED BY OR IS OWNED BY A THIRD PARTY, AS INDICATED IN THE CAPTION LINE, THEN FURTHER PERMISSION MAY BE NEEDED BEFORE ANY FURTHER USE. PLEASE CONTACT WILEY'S PERMISSIONS DEPARTMENT ON PERMISSIONS@WILEY.COM OR USE THE RIGHTSLINK SERVICE BY CLICKING ON THE 'REQUEST PERMISSIONS' LINK ACCOMPANYING THIS ARTICLE. WILEY OR AUTHOR OWNED IMAGES MAY BE USED FOR NON-COMMERCIAL PURPOSES, SUBJECT TO PROPER CITATION OF THE ARTICLE, AUTHOR, AND PUBLISHER. EMBO J, Volume: 38, Issue: 6, First published: 25 February 2019, DOI: (10.15252/embj.2018100024)