Fig. 7.

Cdc34 CTD^prox contacts to Ub(t) and the Ubc core are important for Cdc34 function in cells. a, b Representative western blots (for n = 3) of U2OS cells for Cdc34 and p27 after Cdc34 shRNA KD (a) and subsequent overexpression of WT or mutant Cdc34 (b). c Flow cytometric analysis of U2OS cells treated with control or Cdc34 shRNA then synchronized in G2 phase with nocodazole (middle) and released from synchrony for 12 h (right) with propidium iodide staining to track DNA content. Left, control asynchronous U2OS cell population subjected to the same analysis. Gates indicate percent of live cell population in stages of cell cycle indicated. d Graphical representation of three independent cell cycle experiments as in c and e with the percent of live cells in G1 phase during G2 synchronization (left) and after 12-h release (right) represented by the mean ± SD with individual replicates shown as gray circles. Statistical analysis was conducted using a one-way ANOVA with Bonferroni as Post Hoc test (n = 3), and n.s. indicates non-significant changes while *** indicates a p-value of < 0.001. e Flow cytometric analysis as in c with Cdc34 shRNA KD followed by WT or mutant Cdc34 overexpression as indicated by labels. Source data for all panels are provided as a Source Data file