Extended Data Figure 2 ∣. Imaging Sst⁺ CeA neuron activities with or without learning.

a, Top and middle panels: trial-by-trial licking events for a representative mouse in response to a sound in naïve state (top), and in response to the sound (CS_WA) and water (US_WA) after training in the Pavlovian reward conditioning (middle). Bottom: average licking rate across trials for this mouse after training. b, Top and middle panels: trial-by-trial pupil size change for a representative mouse in response to a sound before training (top), and in response to the sound (CS_SH) and shock (US_SH) after training in the Pavlovian fear conditioning (middle). Bottom: average pupil size change across trials for this mouse after training. c, Quantification of the percentage of Sst⁺ CeA neurons showing inhibitory responses to different stimuli before and after training (n = 4 mice; CS_WA, P = 0.8240, US_WA, P = 0.3534, CS_SH, P = 0.2754; n.s., nonsignificant, paired t-test). d, Pie charts showing the percentage distributions of Sst⁺ CeA neurons according to their response profiles to CS and US after training in reward conditioning (left) or fear conditioning (right). e, Footprints of Sst⁺ CeA neurons in a representative mouse imaged at naïve (left) stage and after training (right). After image alignment, the neurons detected in both sessions are labelled in green. f, Average activity traces of three example tracked neurons in different types of trials at naïve stage (upper two rows) and after training (bottom row). g, Left, the number of neurons detected at different training stages across mice. Right, the percentage of tracked neurons at different training stages across mice (n = 4 mice). h, Quantification of the percentage of tracked neurons among the CS_WA- or US_WA-responsive neurons in trained mice. Note that all the CS_WA-responsive neurons were newly acquired during training; by contrast, 50% of the US_WA-responsive neurons were newly acquired during training, and the other 50% of these neurons existed at the naïve stage. i, The population activities of tracked neurons in one representative mouse in response to CS_WA and US_WA at naïve stage (left) and after training (right). The first three principal components of the population activities are projected onto a 3D space. j, Quantification of the Mahalanobis distance between the vector representing CS_WA responses and that representing US_WA responses across training stages (n = 4 mice; *P = 0.0267, paired t-test). k-p, Tracking Sst⁺ CeA neuron activities across days without learning. k, Four example neurons showed stable responses to water, sucrose, food, or shock on day 1 and day 5. l, Percentage of responsive neurons to each stimulus across mice on day 1 and day 5 (n = 4 mice; water, P = 0.6376; sucrose, P = 0.6139; food, P = 0.5636; shock, *P = 0.0486; n.s., nonsignificant; paired t-test). m, Percentage distributions of neurons excited by different stimuli on day 1, day 3 and day 5 (Fisher’s exact test on the overlaps, water/sucrose: day 1, ****P = 4.28e-13, day 3, ****P < 1e-15, day 5, ****P = 4.09e-11; water/food: day 1, P = 0.3818, day 3, P = 0.1163, day 5, P = 0.1362; water/shock: day 1, P = 0.5178, day 3, P > 0.9999, day 5, P = 0.5527; sucrose/food: day 1, P = 0.2684, day 3, P = 0.2823, day 5, P = 0.8011; food/shock: day 1, P = 0.4434, day 3, P = 0.6465, day 5, P > 0.9999). n, Footprints of Sst⁺ CeA neurons in a representative mouse imaged on day 1 (left) and day 5 (right). After image alignment, the neurons detected in both sessions are labelled in green. o, Neurons were tracked across day 1 and day 5. Each Venn diagram shows the relationship between the neurons excited by a stimulus on day 1 and those excited by the same stimulus on day 5 (Fisher’s exact test on the overlaps: water, ****P = 2.32e-09; sucrose, ****P = 5.36e-06; food, **P = 0.0016; shock, **P = 0.0014). The percentage distributions are calculated based on all the tracked neurons (207 neurons in 4 mice). p, Each pie chart shows, for the neurons excited by a stimulus on day 1, the fractions of these neurons excited by other stimuli on day 5.

Data are presented as mean ± s.e.m. Shaded areas represent s.e.m.