We identified VPS34 as the direct substrate of UBE3C E3 ligase in vivo and in vitro. Next, to analyze the ubiquitin chain on VPS34 that was assembled by UBE3C in vitro.
Ubiquitinated sites on VPS34 identified by LC-MS/MS analysis of VPS34 derived from in vivo ubiquitination assays of UBE3C-transfected or -untransfected 293T cells.