Table 1.

Summary of experiments conducted.

No	Experiment	Species	Sex	Rational	Main figure	Additional outcome
1	Effects of semaglutide on alcohol intake for 8–10 weeks a. Acute, 0.026 mg/kg b. Acute, 0.052 mg/kg c. Repeated, 0.026 mg/kg	Rats	Males Females	High intake over prolonged period of time is important for the manifestation of alcohol use disorder.	1	Influence of semaglutide on food intake (Fig. 2), alcohol preference, water and total fluid intake and body weight (Supplementary Figs. S2, S3 and S5)
2	Effects of semaglutide on relapse-like drinking in the alcohol deprivation model	Rats	Males Females	Abstinence to alcohol causes relapse-like drinking, a behaviour reflecting the inability to abstain from alcohol. This is an important aspect of alcohol use disorder.	1	Influence of semaglutide on alcohol preference, water, total fluid and food intake as well as body weight (Supplementary Fig. S7)
3	Effects of semaglutide treatment on alcohol-induced locomotor stimulation	Mice	Males	Alcohol causes a release of dopamine in NAcS in rodents and humans, an effect associated with an increase locomotor activity, thus reflecting the acute effects of alcohol.	3	Effects on other locomoto activity parameters (Supplementary Fig. S8)
4	Effects of semaglutide treatment on alcohol-induced dopamine release in NAcS, in vivo	Mice	Males	The ability of alcohol to enhance dopamine in NAcS is associated with the rewarding experience in man and is a risk factor for a later alcohol use disorder diagnosis	3	Effects of semaglutide together with alcohol on the release of dopamine metabolites (HVA, DOPAC; Fig. 4). These additional data explore the possibility that semaglutide together with alcohol changes the metabolism of dopamine. This contributes to the identification of underlying molecular underpinnings. Effects on other monoaminergic neurotransmission in NAcS (Supplementary Fig. S11).
5	Effects of semaglutide treatment on alcohol-induced conditioned place preference	Mice	Males	The memory of the alcohol reward is an important aspect of the alcohol use disorder process, a behaviour that can be measured in the memory of conditioned place preference test	Result section
6	Effects of semaglutide treatment on the intake of rewarding foods	Mice	Males	The possibility that semaglutide reduces other types of reward was further explored in feeding experiments, where the intake of rewarding/palatable foods was measured	3	Effects on other feeding parameters at 4-h time point (Supplementary Fig. S9) and at 2-h time point (Supplementary Fig. S10).
7	Effects of alcohol/semaglutide on dopamine-related gene expression	Mice	Males	To explore if semaglutide and alcohol increases the expression of the enzymes metabolizing dopamine (MAOA, MAOB, COMT) in NAc.	4	Gene expression in the VTA (Supplementary Fig. S13).
8	Effects of semaglutide on ex vivo dopaminergic neurotransmission in NAcS and NAcC from alcohol drinking rats of both sexes.	Rats	Males Females	To further explore the possibility that semaglutide together with alcohol changes dopamine metabolism	Supplementary Fig. S12	Additional ex vivo data from other brain areas (Supplementary Fig. S13 and Supplementary Table S1–S2)
9	Effects of semaglutide on other dopamine driven behaviours, alcohol naïve	Rats	Males Females	To explore the possibility that semaglutide affects exploratory behaviours and novelty seeking.	Supplementary Fig. S12
10	Detection of fluorescently labelled semaglutide in NACS of alcohol drinking rats	Rats	Males Females	As semaglutide increases the metabolism of dopamine in NAcS when alcohol is onboard, we hypothesise that fluorescently labelled semaglutide may be preset and exert its effects locally in NAc.	3
11	Effects of semaglutide on cAMP in NAc, alcohol naïve	Mice	Males Females	To understand the effects of semaglutide within the NAc on cAMP, a key downstream signalling molecule of GLP-1R was measured in the NAc	Supplementary Fig. S14	cAMP in amygdala, VTA, NTS (Supplementary Fig. S14)
12	Effects of semaglutide on activity and anxiety-like behaviours	Rats	Males Females	Activity and anxiety are tentative confounding factors to the semaglutide reduced alcohol intake, a possibility explored in locomotor activity and elevated plus maze experiments	Supplementary Fig. S15

Summary of all experiments conducted. Description of species and sex used, the rational for conducting the experiments, description figure/supplementary material where the results can be found, and additional outcome obtained (besides the main findings). Nucleus accumbens shell (NAcS) and core (NAcC), ventral tegmental area (VTA), nucleus tractus solitarius (NTS), glucagon-like peptide-1 (GLP-1R).