Extended Data Figure 6:

Gel panels in this figure are representative of two independent experiments; n = 2. a, A truncated form of the human deubiquitinating enzyme (DUB) TRABID (residues 245-697, named Tra) was used to reduce the level of K29-linked polyubiquitination in HeLa cells. Domain diagrams of full-length TRABID and the Tra construct are shown. b, Western blots of K63-linked ubiquitin and total ubiquitin in HeLa cells transfected with the Tra construct or the empty vector. c, Cell cycle analysis of HeLa cells transfected with either the Tra or the Tra-CIM construct by flow cytometry. cMyc and K29-linked polyubiquitin channels are shown. Representative flow cytometry graphs from three biological replicates are shown. d, Cell cycle analysis of HeLa cells transfected with either the Tra construct or the empty vector by flow cytometry. cMyc and K29-linked polyubiquitin channels are shown. Note that the cMyc tag was also expressed in cells transfected with the empty vector. Representative flow cytometry graphs from two biological replicates are shown. e, Cell cycle analysis of normal HeLa cells by flow cytometry. f, Cell cycle analysis of Hela cells transfected Tra construct for 27 h using flow cytometry. Representative flow cytometry graphs from three biological replicates are shown (a total of 5 injections). Highlighted areas were subjected to cell cycle analysis.