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. 2009 Jul 13;29(18):4959–4970. doi: 10.1128/MCB.00562-09

FIG. 3.

FIG. 3.

Degradation of Rep2 is mediated through APC/C but not the SCF complex. (A) Rep2 is stabilized in lid1-6 cells. Western blot analysis of Rep2-HA levels in lid1-6 cells at various time points after addition of CHX is shown. Prior to CHX addition, cells were grown at 36°C for 4 h to inactivate lid1-6. (B) Rep2 is unstable in skp1-4A cells. Western blot analysis of Rep2-HA levels in skp1-4A cells after addition of CHX is shown. Prior to CHX addition, cells were grown at 36°C for 4 h to inactivate skp1-4A. (C) Inactivation of Lid1 but not Skp1 diminishes the polyubiquitination of Rep2. In vivo ubiquitin assays show the Ni-NTA bead-enriched polyubiquitin-conjugated proteins using antiubiquitin antibodies (left panel) and after rehybridization with anti-HA antibodies (right panel). The position of slowly migrating polyubiquitinated Rep2 (Ubi-Rep2) is indicated. M, molecular mass.