Supplementary Figure 8: Chronic Oxt^PVH inactivation increases water intake following dehydration, salt depletion, mannitol and PEG

(a) Injection of AAV-DIO-GFP:TetTox in Oxt^PVH neurons. (b), Representative images of GFP expression in Oxt^PVH neurons in GFP:TetTox- vs control YFP-injected mice; 3V, third ventricle; (n = 6 TetTox, 8 YFP) scale bar 100 µm. (c) Chronic Oxt^PVH inactivation does not significantly alter baseline saline preference at different concentrations (0.075, 0.3, 0.5 M) (n = 6 TetTox, 8 YFP; unpaired 2-tailed Student’s t-test; 0.075 M NaCl: interaction t(12) = 0.2346; p = 0.8184; 0.3 M NaCl: interaction t(12) = 0.2509; p = 0.8061; 0.5 M NaCl: interaction t(12) = 0.6848; p = 0.5065), but (d) but significantly increases water following 24-h dehydration (n = 6 TetTox, 8 YFP; 2-way RM ANOVA; NaCl: interaction F(8,96) = 1.098; p = 0.3712; water: interaction F(8,96) = 4.073; p = 0.0003), (e) 48-h salt depletion (n = 6 TetTox, 8 YFP; 2-way RM ANOVA; NaCl: interaction F(8,96) = 0.7341; p = 0.6612; water: interaction F(8,96) = 6.866; p < 0.0001), (f) demonstrates a trend to increase water following 0.5 M saline ip (n = 6 TetTox, 8 YFP; 2-way RM ANOVA; NaCl: interaction F(4,48) = 0.1292; p = 0.9711; water: interaction F(4,48) = 2.183; p = 0.0850), and (g) increases water following 1 M mannitol ip (n = 6 TetTox, 8 YFP; 2-way RM ANOVA; NaCl: interaction F(4,48) = 0.8378; p  = 0.5080; water: interaction F(4,48) = 3.628; p = 0.0116), and (h) and 30% PEG sc (n = 6 TetTox, 8 YFP; 2-way RM ANOVA; NaCl: interaction F(4,48) = 1.296; p = 0.2848; water: interaction F(4,48) = 3.617; p = 0.0118). Data are expressed as mean ± s.e.m. ****p < 0.0001; **p < 0.01; *p < 0.05; φ p = 0.085. See Supplementary Information for statistical analyses