FBN rats were fed normal (0.9%) or 8% dietary salt for 7 days. The animals were euthanized following blood pressure measurement on day 7 and the kidneys were removed, decapsulated, and crude membrane homogenates were prepared as described in methods. Expression of (A) NHERF1 (top panel); D1R (middle panel); and NKA α1 (bottom panel). One gel was used, and the nitrocellulose paper was cut below the 100 KDa marker (for NKA α1), at 60 KDa (for D1R) and the bottom portion below 60 KDa was used for NHERF1 together with Actin (B) NKCC2; and (C) NCC was determined by western blot. A representative western blot from one male and one female rat groups is shown. To determine equal loading the nitrocellulose membrane was simultaneously blotted on the same blot for actin using an HRP-actin antibody together with HRP-secondary antibodies. Each data point represents AU (ratio of specific protein to actin) from a single animal (red=normal salt-diet; blue=8% salt diet) and the horizontal lines are data as mean±SD from 9 animals in each group (4 males and 5 females). *** indicates p < 0.001 and * indicates p < 0.05 from normal salt fed rats, by two-way ANOVA followed by Bonferroni post-hoc test.