In female rats, E₂ may suppress the depolarization of E_GABA in AVP neurons associated with DOCA-salt treatment by inhibiting the up-regulation of NKCC1 and down-regulation of KCC2. (A–C) Effects of the NKCC1 blocker bumetanide (10 μmole/L) and the KCC2 blocker VU0463271 (5 μmole/L) on E_GABA in AVP neurons recorded in the SON slices from DOCA-salt-treated Sham, OVX, and OVX+E₂ rats. The bar charts are the E_GABA values of these cells. The symbols connected by lines indicate data from the same cells, and the same kind of symbols indicate that they were from the same rat. n: number of rats used for the preparation of the SON slices. **P < 0.01, ***P < 0.001 compared with the value before drug application (one-way repeated measures ANOVA followed by pairwise comparison with the Holm–Sidak procedure). (D) NKCC1 (145∼205 KDa), KCC2 (140∼270 KDa), and β-actin (∼40 KDa) bands recognized by western blotting of the SON tissues from DOCA-salt-treated Sham, OVX and OVX+E₂ rats. (E and F) Bar charts showing the relative levels of NKCC1 and KCC2 in the SON of these rats. For each experiment, values were normalized to the mean value of the samples collected from Sham rats. *P < 0.05 (Kruskal–Wallis one-way ANOVA on ranks followed by pairwise comparison with Student–Newman–Keuls method). n: number of samples.