Tissue extracts (30 μg of protein) prepared from two individual SGK1 -/- mice and two individual SGK1 +/+ mice were subjected to SDS/PAGE, transferred to ImmobilonP membranes and immunoblotted with anti-GST-NDRG1 (upper panel) or anti-NDRG1 phospho Thr x 3 (lower panel) at a concentration of 1 μg/ml. Binding of the primary antibody was detected using rabbit peroxidase conjugated anti- sheep IgG antibody (1 in 10, 000 dilution, Pierce) followed by enhanced chemiluminescence (ECL, Amersham).