Extended Data Fig. 5: Control experiments for optogenetic manipulation.

a–g, Optogenetic inhibition of Sst⁺ CeA neurons during US presentation in well-trained mice does not impair performance. a, A representative ArchT mouse (left) and GFP mouse (right) used in Fig. 4 was given additional 8 sessions of training in the go/no-go task in the absence of laser stimulation. The licking events, sorted according to trial types, for the two mice in session 8 (S8) of the additional training, in which they have reached similar levels of performance and anticipatory licking. b, All the mice used in Fig. 4 were given additional 8 sessions of training in the go/no-go task in the absence of laser stimulation. Their performance and anticipatory licking rate in different sessions and trial types were quantified (ArchT group, n = 6 mice, GFP group, n = 5 mice; last session, n.s., nonsignificant, P > 0.05; t-test). c & d, The same mice as in b were tested in another 3 sessions, in which the laser stimulation was delivered to the CeA immediately following the onset of US delivery in each trial. c, Behavior of a representative ArchT mouse (left) and GFP mouse (right) in the 3^rd of the test sessions. The licking events were sorted according to trial types. Dashed lines indicate the onset of CS and US. The green shaded area indicates the time window in a trial when the laser was turned on. d, Behavior of all the mice across sessions during the test. Their performance and anticipatory licking rate in different sessions and trial types were quantified (n.s., P > 0.05; two-way ANOVA). e, Confocal histological images of coronal brain sections from a representative mouse used for the experiments, showing ArchT-GFP expression in Sst⁺ CeA neurons and the locations of optical fiber implantation. f, Schematics showing the placement of fiber implants in the ArchT mice (n = 6 mice) used for the experiments. g, Confocal histological images of a coronal brain section from a representative mouse, showing ArchT-GFP expression (left) and the expression of Sst recognized by an antibody (middle). Almost all ArchT-GFP⁺ cells are also Sst⁺ (right). h–o, Optogenetic inhibition of Sst⁺ CeA neurons during CS presentation in well-trained mice does not impair performance. h & i, Behavior of well-trained ArchT mice in the reward-only task. h, The licking events of a representative mouse sorted according to laser and no-laser trials. Dashed lines indicate the onset of CS and US. The green shaded area (2 s) indicates the time window in a trial when the laser was turned on (50% of trials). i, The performance (left) and anticipatory licking rate (right) of all the mice in laser and no-laser trials (n = 7 mice, n.s., nonsignificant, P > 0.05; paired t-test). j & k, Behavior of the same mice in i after being well trained in the go/no-go task. j, The licking events of a representative mouse sorted according to laser and no-laser trials in the go trials. Dashed lines indicate the onset of CS and US. The green shaded area (2 s) indicates the time window in a trial when the laser was turned on (50% of go trials). k, The performance (left) and anticipatory licking rate (right) of all the mice in laser and no-laser trials (n.s., P > 0.05; paired t-test). l & m, Same as j & k, except that the no-go trials are used for the presentation (l) and analyses (m) (n.s., P > 0.05; paired t-test). n, Histological images of coronal brain sections from a representative mouse used for the experiments, showing ArchT-GFP expression in Sst⁺ CeA neurons and the locations of optical fiber implantation. o, Schematics showing the placement of fiber implants in the ArchT mice (n = 7 mice) used for the experiments. p–t, Optogenetic inhibition of Sst⁺ CeA neurons does not induce aversion or preference and has no effect on movements. p, Heat-maps for the activity of a representative ArchT mouse at baseline (top), or in a situation whereby entering the left (middle) or right (bottom) side of the chamber triggered photo-inhibition of Sst⁺ CeA neurons (i.e., the real-time place preference/aversion (RTPP/RTPA) test). q, Quantification of the behavior as shown in a, for mice in which Sst⁺ CeA neurons expressed ArchT (n = 6 mice, top) or GFP (n = 7 mice, bottom) (n.s., nonsignificant), P > 0.05, one-way ANOVA. r, Quantification of moving velocity (top) and distance (bottom) for the ArchT mice in q in the RTPP/RTPA test (n.s., P > 0.05, one-way ANOVA). s & t, Experiment with the continuous licking task. s, A schematic of the setup for the task. t, Quantification of the effect of laser stimulation on licking rate in ArchT mice and GFP mice (n.s., P > 0.05, one-way ANOVA). u–w, Optogenetic inhibition of Sst^CeA→DA projections in the go/no-go task after mice learned the reward task. The same mice used in Fig. 5b were further trained in the absence of laser stimulation such that the ArchT group and GFP group reached similar performance in the reward-only task. The two groups were subsequently trained in the go/no-go task, during which a green light (3 s) was delivered into the CeA immediately after the onset of US presentation in each trial throughout the training. u, Top left, hit rate across training sessions (ArchT group, n = 10 mice, GFP group, n = 11 mice; F(1,19) = 0.1965, n.s., nonsignificant, P = 0.6626; two-way ANOVA). Top right, licking rate following CS onset in go trials across training sessions (F(1,19) = 0.4038, n.s., P = 0.5327; two-way ANOVA). Bottom left, false alarm rate across training sessions (F(1,19) = 0.1985, n.s., P = 0.6610; two-way ANOVA). Bottom right, licking rate following CS onset in no-go trials across training sessions (F(1,19) = 0.7804, n.s., P = 0.388; two-way ANOVA). v, Histological images of coronal brain sections from a representative mouse used for the experiments, showing ArchT-GFP expression in Sst⁺ CeA neurons (top) and the locations of optical fiber implantation in the SNc (bottom). w, Schematics showing the placement of fiber implants in the SNc of the ArchT mice used for the experiment. Data are presented as mean ± s.e.m.