FIGURE 1.

HUWE1 interacts with USP7 but does not target the protein for degradation. A, HEK 293T cells were transfected with empty vector (EV) or Avi-tagged CD HUWE1 and subsequently treated with MG132 prior to lysis. Lysates were incubated with streptavidin resin, resolved by SDS-PAGE, and stained with Coomassie Blue. B, scatter plot representing proteins identified by LC-MS/MS following pull-down of Avi-tagged CD HUWE1. Plotted on the y axis are the log₁₀ ratios of proteins from cells expressing HUWE1 compared with a vector control sample. On the x axis is the p value indicating statistical significance of each protein. C, HEK 293T cells were transfected with FLAG-tagged candidate HUWE1-interacting partners or empty vector (EV) for 24 h. Input whole cell lysates were subjected to immunoprecipitation (IP) with FLAG affinity resin, and samples were analyzed by immunoblotting. D, immunoblot analysis of USP7 from HEK 293T cells transfected with a non-targeting (NT) control siRNA or HUWE1 siRNAs for 72 h. E, HEK 293T cells were transfected with a non-targeting (NT) control or USP7 siRNAs for 72 h and analyzed by immunoblotting.