The animals were then food deprived for 56 h (IACUC approved), a time length previously shown to maximize food hoarding [4] and [18]. Before access to food was returned at light offset, half of the animals received an injection of PYY(3-36) (0.1 nmol in 200 nl), the active form of the peptide for satiation [52], into the GSK-3 cancer Arc and the other half received the saline vehicle. Wheel revolutions,

food foraging, food intake, and food hoarding were measured at 1, 2, 4, 24 h and each day post-injection until all animals returned to pre-injection levels. After the animals returned to behavioral baseline, brain tissue was collected to verify cannula location (Fig. 1; 69 hits and 11 misses or removed their cannula; final group sizes: PYY(3-36): BW: n = 12, FW: n = 11, and 10REV: n = 13 and vehicle: BW: n = 9, FW: n = 11, and 10REV: n = 13). Raw data from Experiment 1 were transformed for each individual into percent change from vehicle before

statistical analyses using the formula: [((X-Vehicle)/Vehicle) × 100], where “X” equals the value measured in response to the dose of BIIE0246 FLT3 inhibitor and “Vehicle” equals the value measured for that individual after vehicle injection. No statistical comparisons were made among the time intervals because the intervals were of unequal duration. No statistical comparisons are reported across test days in this counterbalanced-within subject design, as repeated measures two-way ANOVA (foraging treatment × Arc-injection) showed no effect of injection order. The data were analyzed using a two-way ANOVA (foraging treatment × Arc-injection; 3 × 4). For Experiment Niclosamide 2, data were not transformed into percent change from

vehicle, because animals only were food deprived once and therefore could not serve as their own control, and the absolute values were analyzed using a two-way ANOVA (foraging treatment × Arc-injection; 3 × 2) within each individual time point for the same reason as above. All statistical analyses were performed using NCSS (version 2007, Kaysville, UT). Exact probabilities and test values were omitted for simplicity and clarity of presentation. Differences were considered statistically significant if P < 0.05. Tukey-Kramer Multiple Comparison Tests were used for post hoc tests when appropriate. Misplaced cannulae were not included in the final statistical comparisons. Wheel running. At each time interval, Arc injection of BIIE0246 did not significantly stimulate wheel running activity compared to vehicle injection at any of the three doses tested (0.1, 1.0 and 5.0 nmol; Fig. 2A). The lack of wheel running increase in the FW group, where food delivery was not contingent upon wheel running, suggests that there was not non-specific stimulation of locomotor activity. Food foraging.