Fig. 7: UCHL1 and UCHL5 are substrates of UBE2W.
From: Antibody toolkit reveals N-terminally ubiquitinated substrates of UBE2W
a In vitro ubiquitination assays were performed on lysine-less (K0) catalytically inactive UCHL1C90S (top) and UCHL5C88S (bottom). Two-hour reactions were carried out in the absence of UBE2W (lane 1 and 2) and the presence of UBE2W (lane 3–5). All reactions were incubated with E1, E3 (RNF4), ATP/MgCl2, and with (lane 2) or without (lane 5) ubiquitin. Results are representative of three independent experiments. b Western blots of doxycycline-inducible UBE2W/RNF4 and UBE2WW144E/RNF4 HEK293 cells at 24 h post doxycycline treatment. Endogenous UCHL1 expression was analyzed with an anti-UCHL1 antibody. Results are representative of three independent experiments. Source data are provided as a Source Data file.
