Fig. 3.

ENaC and Sgk1 are regulated by aldosterone in IMCD3 cells. A and B: total RNA was isolated from IMCD3 cells cultured in DMEM plus 10% charcoal-stripped FBS for at least 50 h and then treated with vehicle (ethanol, −Aldo) or 1 μM aldosterone (+Aldo) for 24 h and analyzed by real-time RT-quantitative PCR for expression of ENaC subunits (A), and ENaC transcriptional regulators Sgk1 and MR (B). The mRNA level of each gene was first normalized against β-actin mRNA in the same sample and set to 1 in the vehicle-treated cells. *P < 0.05 vs. −Aldo; n = 3. C: as in A, whole cell lysates were analyzed by immunoblotting with antibodies specific for the proteins as indicated. For each protein, the relative band intensity in vehicle-treated cells was set to 100 for comparison. MR, mineralocorticoid receptor. *P < 0.05 vs. −Aldo; n = 3.