Fig. 4: NI^RLN3 responses to CS presentation.

a Schematic for NI^RLN3 GCaMP recordings (left) with representative image showing GCaMP transduction and implantation site of optical fiber (right). Scale bar, 100 µm. b–d NI^RLN3 fiber photometry signals in GFP- or GCaMP7s-injected mice responding to conditioned stimulus (CS) under fed (b) and fasted (c) states. The normalized change in fluorescence (ΔF/F) was calculated. b, c Black dashed line indicates CS onset (time = 0); solid lines indicate average fluorescence traces, while shadings represent 95% confidence interval; gray frames show 5-s windows post-CS onset, which are used for calculations of area under the curve (AUC). b–d n = 14 trials/7 mice/group. e Schematic for NI^RLN3 GCaMP recording accompanying inhibition of LH inputs (left), and representative image demonstrating GCaMP7s expression and sites of optical fiber and cannula (right). Scale bar, 100 µm. f, g NI^RLN3 fiber photometry signals in fasted mCherry- or hM4Di-injected mice responding to CS after the infusion of clozapine-N-oxide (CNO) into the NI. f Black dashed line indicates CS onset (time = 0); solid lines indicate average fluorescence traces, while shadings represent 95% confidence interval; gray frames show 5-s windows post-CS onset, which are used for calculations of area under the curve (AUC). f, g n = 10 trials/5 mCherry mice, 12 trials/6 hM4Di mice. Data were presented as the mean ± SEM; *p < 0.05, **p < 0.01. Statistical significance was assessed using two-way repeated measures (RM) ANOVA with uncorrected Fisher’s LSD multiple-comparisons test (d) or two-sided unpaired t-test (g). Source data and exact p value are provided as a Source Data file.