Mouse anti ezrin

The antibodies used were: rabbit anti-SGK1 antibody (Sigma); mouse anti-CFTR C-terminus antibody (clone 24-1; R&D systems, Minneapolis, MN); mouse anti-CFTR antibody (clone 596; University of North Carolina Cystic Fibrosis Center, Chapel Hill, NC); mouse anti-ezrin, mouse anti-Rab5a and mouse anti-EEA1 antibodies (BD Biosciences, San Jose, CA); rabbit anti-Rab 11a (Life Technologies Corp); mouse anti-EGFR (Santa Cruz Biotechnology, Inc, Santa Cruz, CA); horseradish peroxidase-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (Bio-Rad, Hercules, CA). SGK1-S442D, a constitutively active form of human SGK1 and human SGK1-K127N, an inactive form of human SGK1, were constructed as described [15] . The SGK inhibitor GSK 650394 was obtained from Tocris (purchased through R&D Systems, Minneapolis, MN). All antibodies and reagents were used at the concentrations recommended by the manufacturers.

Unless noted, all chemicals were purchased from Sigma-Aldrich. The following antibodies were used: Mouse anti-Ezrin (BD Biosciences), rabbit anti-NHERF2 (Sigma-Aldrich), rabbit anti-EBP50 and anti-HA (Abcam), rabbit anti-Actin (Millipore). Secondary antibodies were raised in goat and purchased from Life Technologies and Jackson ImmunoResearch Laboratories. Phalloidin-568 (Life Technologies) was made up at 2 mg/ml in methanol and used at a final concentration of 1 μg/ml. During experiments, cells were maintained in a modified Ringer solution containing (mM): 130 NaCl, 5.1 KCl, 1.2 CaCl₂, 1.2 MgCl₂, 10 mM HEPES, 10 glucose, pH 7.4.