Supplementary Figure 2: Changes in pupil area and locomotion across states do not account for observed changes in BA activity.

a, Mean pupil area in the 2 s preceding a cue during hungry and sated states (normalized to maximum pupil area during the entire session across both states). Mean ± s.e.m. across 15 sessions from 7 mice. ** p = 0.0012, two-sided Wilcoxon sign-rank. b, Mean speed of locomotion in the 2 s preceding a cue during hungry and sated states (normalized to maximum locomotion during the entire session across both states). Mean ± s.e.m. across 15 sessions from 7 mice. *** p = 0.0006, two-sided Wilcoxon sign-rank. c, Histograms of pupil areas in the 2 s preceding onset of each cue during hungry and sated states from an example session before (left) and after (right) matching trials for pupil area (see Methods). Note that following matching of trials, distributions of pupil areas are now completely overlapping. Insets show moments of small (left) and large (right) pupil diameter (bright ellipse against a dark background, as infra-red laser light emitted from the pupil was used for tracking, see Methods). d, Histograms of relative locomotion speed in the 2 s preceding cues during hungry and sated states from an example session before (left) and after (right) matching trials for locomotion. e, Comparison of cue responses (mean ± s.e.m.) of BA neurons across hungry and sated states before and after matching for pupil areas. The finding of attenuated cue responses following satiation persisted even when matching pupil area distributions across states. Sample size for RC: n = 66 activated neurons and n = 58 suppressed neurons; for AC-Av: n = 13 activated neurons and n = 26 suppressed neurons; for NC: n = 21 activated neurons and n = 25 suppressed neurons. Original AC-Av activated response magnitude, hungry vs. sated: ** p = 0.0089. Pupil-matched AC-Av activated response magnitude hungry vs. sated: ** p = 0.0061. For every other comparison, *** p < 0.0001, two-sided Wilcoxon sign-rank. Mean response magnitudes were analyzed separately for neurons that were significantly activated (red) or suppressed (blue) by cue presentation. f, Comparison of cue responses (mean ± s.e.m.) of BA neurons across hungry and sated states before and after matching for locomotion. The finding of attenuated cue responses following satiation persisted even when matching locomotion distributions across states. Sample sizes are same as in panel e. Original AC-Av activated response magnitude, hungry vs. sated: ** p = 0.0012. Locomotion-matched AC-Av activated response magnitude hungry vs. sated: * p = 0.019. Original NC activated response magnitude hungry vs. sated: *** p < 0.0001. Locomotion-matched NC activated response magnitude, hungry vs. sated: ** p = 0.0026. Original NC suppressed response magnitude, hungry vs. sated: *** p < 0.0001. Locomotion-matched NC suppressed response magnitude, hungry vs. sated: * p = 0.027. For every other comparison, *** p < 0.0001, two-sided Wilcoxon sign-rank.