Extended Data Figure 3. Regulation of SFO^Nos1 neurons by homeostatic signals.

a, Recordings from SFO^Nos1::GCaMP6s mice as they explored a behavioral chamber without access to food or water revealed dynamic fluctuations in fluorescence around a stable baseline (1 of 8 mice); these fluctuations were absent from recordings from SFO^Nos1::GFP mice (1 of 3 mice). b, Quantification of response to peripheral injection of NaCl (averaged traces in Fig. 1b; **P < 0.01, ****P < 0.0001, two-way repeated-measures ANOVA, n = 5 mice). c, Time constant (τ) of rising and falling phases of response to peripheral injection of NaCl (n.s., two-way repeated-measures ANOVA, n = 5 mice). d, Representative recordings for five mice showing response to peripheral injection of NaCl or vehicle. e, SFO^Nos1 neurons are activated by peripheral injection of angiotensin in a dose-dependent manner (n = 3 mice). f, Quantification (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, two-way repeated-measures ANOVA, n = 3 mice). g, The AT₁R antagonist losartan abolished the response to peripheral injection of angiotensin (n = 3 mice). h, Quantification (**P < 0.01, two-way repeated-measures ANOVA, n = 3 mice). i, Schematic illustrating expected observations if activation of SFO^Nos1 neurons in response to peripheral injection PEG/isoproterenol and NaCl is angiotensin-dependent. j, Angiotensin blockers abolished the response to peripheral injection of PEG (quantification in Fig. 1f; n = 5 mice). k, Angiotensin blockers abolished the response to peripheral injection of isoproterenol (quantification in Fig. 1h; n = 5 mice). l, Angiotensin blockers did not abolish the response to peripheral injection of NaCl (quantification in Fig. 1c; n = 5 mice). m, Schematic illustrating expected observations if activation of SFO^Nos1 neurons in response to peripheral injection NaCl is sodium-sensory or osmo-sensory. n, SFO^Nos1 neurons are similarly activated by peripheral injection of equimolar mannitol and NaCl (quantification in Fig. 1d; n = 5 mice). Values are mean ± s.e.m. (error bars or shaded area).