Figure 2.
The ERAD of ENaC subunits is dependent on both the Hrd1 and Doa10 ubiquitin ligases. Cycloheximide chase reactions were performed as described in Materials and Methods in HRD1/DOA10 (●), hrd1Δ (▴), doa10Δ (▵), or hrd1Δdoa10Δ (○) yeast strains (Pagant et al., 2007) expressing C-terminally HA-tagged α-, β-, or γ-ENaC. Chase reactions were performed at 37°C, lysates were immunoblotted with anti-HA (ENaC) or with anti-Sec61 (as a loading control). Data represent the means of 4–6 experiments, ±SEM.