Extended Data Fig. 9: DREADD manipulation of the ACC selectively influences competitive effort but not motivation or reward-seeking behaviour.

a, b, Overlay of viral expression areas for a. hM3D(Gq)-mCherry across 13 animals and for b. hM4D(Gi)-mCherry across 12 animals. cg1/cg2=cingulate areas 1/2; prL=prelimbic cortex; iL=infralimbic cortex. c, Novel intruder assay for social aggression. Although the animals displayed an increase in social inspection behaviours following CNO compared saline (Left, *Z=1.77, p=0.038, one-sided rank sum), they displayed no change in attack or defensive behaviours (Right, p>0.3, two-sided Rank-sum). N=52 trials across n=15 animals. d, The likelihood of competitive success was higher for ACC excitation (Left, n=13, compared to saline) in trials where they were of lower relative rank (relative rank 3 or 4) than their competitors (*t₍₁₂₎=2.63, p=0.011; Paired t-test), but not when they were higher in relative rank (relative rank 1 or 2) than their competitors (t₍₁₂₎=-0.24, p=0.59; Paired t-test). Right, The likelihood of competitive success was decreased for ACC inhibition (n=12, compared to saline) in trials where they were of higher relative rank (relative rank 1 or 2) than their competitors (**t₍₁₁₎=-2.11, p=0.028; Paired t-test), but not when they were of lower relative rank (relative rank 3 or 4) than their competitors (t₍₁₁₎=-0.25, p=0.4; Paired t-test; Fig. 4e). e, Animals with ACC excitation reached the reward zone faster (n=13, *F_(1,109)=3.98,p=0.049) in high compared to low reward trials (**F_(1,109)=4.33,p=0.04) and when starting in the near compared to far staging areas (***F_(1,109)=27,p=1.04x10^-17), but there were no interactions between any of the conditions (F_{drug:reward(1,109)}=0.23, p=0.63; F_{drug:stagingarea(1,109)}=0.66, p=0.42; F_{reward:stagingarea(1,109)}=0.52, p=0.47; three-way ANOVA). f, Animals with ACC inhibition reached the reward zone faster in high compared to low reward trials (n=12, *F_(1,95)=5.1, p=0.026) and when starting in the near compared to far staging areas (**F_(1,95)=4.1,p=0.046), but there were no interactions between any of the conditions (F_{drug:reward(1,95)}=0.13, p=0.72; F_{drug:stagingarea(1,95)}=0.043, p=0.84; F_{reward:stagingarea(1,95)}=0.51, p=0.33; three-way ANOVA). g, Mice foraging alone with inanimate totems. Left, There was no difference in latency to reach reward for either ACC excitation (n=13 mice, t₍₁₂₎=-0.058, p=0.96; Paired t-test) or inhibition (n=12 mice, t₍₁₁₎=0.051, p=0.96; Paired t-test). Right, There was no difference in average path error to reaching the reward zone for either ACC excitation (t₍₁₂₎=-0.79, p=0.44; Paired t-test) or inhibition (t₍₁₁₎=0.47, p=0.65; Paired t-test). h, Mice moving a mass of variable weight to receive reward. Left, there was no difference in the latency to reach the reward zone based on differences in weight (low vs. high) for either ACC excitation (n=14 sessions across n=13 mice, F_(2,77)=0.07, p=0.93; Two-way ANOVA) or inhibition (n=12 sessions across n=12 mice, F_(2,67)=0.58, p=0.56; Two-way ANOVA). Right, There was no difference in the latency to push past the weight to reach the reward zone for either ACC excitation (F_(2,77)=0.85, p=0.8; Two-way ANOVA) or inhibition (F_(2,67)=0.01, p=0.99; Two-way ANOVA). Error bars denote mean ± s.e.m. Dots represent session averages. N=6 mice for Exc group and n=6 mice for Inh group.

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