ENaC expressed in the apical membrane of epithelial cells plays a critical role in homeostasis of fluid content and blood pressure. Na+ transport via ENaC depends on not only the open probability of the channel, but also the number of channel expressed on the apical membrane. ENaC is cleaved by aldosterone‐induced proteases, resulting in an increase in open probability. The lifetime of ENaC at the apical membrane is regulated by ubiquitin ligase (Nedd4‐2). To elucidate roles of raft domain in the regulation of ENaC activation, we quantified the abundance and localization of ENaC at the plasma membrane with/without aldosterone stimulation in renal epithelial cells. Aldosterone stimulated expression of both full‐length and cleaved ENaCs; i.e., the full‐length ENaC was expressed in both cytoplasm and plasma membrane (raft domain), whereas a large portion of the cleaved ENaC was observed at the non‐raft plasma membrane. Depletion of cholesterol from the raft domain diminished expression of cleaved ENaC not only in raft but also non‐raft domain by aldosterone stimulation. Aldosterone also increased expression of Nedd4‐2 and SGK1 in raft domain. Together, these results suggest that the ubiquitination‐mediated regulation of ENaC would occur in raft domain. Our findings demonstrate novel roles of the lipid raft domain on the regulation of ENaC activity/trafficking.