From the total of 85 animals KRX-0401 tested, 49 rats had bilateral injections correctly placed into LPBN. Misplaced injections that reached tissue surrounding the LPBN were located mainly dorsal or ventral to LPBN and some of them medial to the LPBN. Results from rats with misplaced injections of suramin or α,β-methylene

ATP were also analyzed and reported to confirm the specificity of the LPBN as the site of injections that produce the effects on sodium intake. ANOVA showed significant differences on sodium depletion-induced 1.8% NaCl intake comparing rats treated with bilateral injections of different doses of α,β-methylene ATP or saline into the LPBN [F(3,24) = 13.39; p < 0.001] ( Fig. 2A). Bilateral injections of the highest dose of α,β-methylene ATP (4.0 nmol/0.2 μl each site) into the LPBN increased sodium depletion-induced 1.8% NaCl intake from 15 to 120 min of the Cilengitide test with p values ranging from p < 0.01 at 15 min to p < 0.001 from 45 to 120 min (Newman–Keuls post hoc test) ( Fig. 2A). The injections of the intermediate dose of α,β-methylene ATP (2.0 nmol/0.2 μl each site) into the LPBN increased sodium depletion-induced 1.8% NaCl intake from 45 to 120 min of test with p values ranging from p < 0.005 (at 45 min) to p < 0.001 (from 60 to 120 min, Newman–Keuls post hoc test) ( Fig. 2A). Bilateral

injections of the lowest dose of α,β-methylene ATP (1.0 nmol/0.2 μl each site) into the LPBN did not change sodium depletion-induced

1.8% NaCl intake ( Fig. 2A). Injections of α,β-methylene ATP (1.0, 2.0 and 4.0 nmol/0.2 μl) produced no effect on water intake (3.5 ± 1.1, 1.3 ± 0.8, 4.4 ± 1.2 ml/120 min, respectively, vs. vehicle: 3.3 ± 1.4 ml/120 min) [F(3,24) = 1.56; p > 0.05] ( Fig. 2B). Bilateral injections of α,β-methylene ATP (2.0 nmol/0.2 μl each site) into the LPBN in sodium replete rats produced no effect on 1.8% NaCl (1.3 ± 0.9 vs. saline: 0.8 ± 0.4 ml/120 min, Amrubicin n = 6) or water intake (0.2 ± 0.1 vs. saline: 0.6 ± 0.3 ml/120 min). ANOVA showed significant differences on sodium depletion-induced 1.8% NaCl intake comparing rats treated with bilateral injections of α,β-methylene ATP (2.0 nmol/0.2 μl each site) or saline after pretreatment with PPADS (4 nmol/0.2 μl) or saline into the LPBN [F(3,27) = 10.97; p < 0.001] ( Fig. 3A). Bilateral injections of α,β-methylene ATP (2.0 nmol/0.2 μl each site) after pretreatment with saline into the LPBN increased sodium depletion-induced 1.8% NaCl intake from 30 to 120 min of the test with p values ranging from p < 0.05 at 30 min to p < 0.005 at 90 and 120 min (Newman–Keuls post hoc test) ( Fig. 3A). Bilateral injections of PPADS (4 nmol/0.2 μl) + saline into the LPBN did not change 1.8% NaCl intake (p ≥ 0.1 at any of the times studied, Newman–Keuls post hoc test) ( Fig. 3A).