Fig. 3: Chronic LA-Agonist treatment has similar plasma metabolomics as muGIPR-Ab treatment, associated with agonist-induced receptor desensitization.

a–e Global metabolomics analysis of plasma samples for vehicle, LA-Agonist, and muGIPR-Ab treated mice (presented in Fig. 2a–k). Significantly different number of metabolites between a vehicle vs. muGIPR-Ab, b vehicle vs. LA-Agonist, and c muGIPR-Ab vs. LA-Agonist. n = 8 mice/group; Welch’s two-sample t-test p ≤ 0.05 and Storey method for correcting multiple comparisons q-value ≤ 0.2. d Principal component analysis (PCA) of the plasma metabolites from global metabolomics analysis. e Significantly enriched pathways between vehicle vs. muGIPR-Ab and vehicle vs. LA-Agonist treatments. f–h Plasma metabolites f cAMP, g corticosterone, and h glycerol. Data are scaled such that the median value measured across all samples was set to 1.0 ± SEM, n = 8 mice/group; Welch’s two-sample t-test, f Vehicle vs. LA-Agonist *p = 0.0101 and Vehicle vs. muGIPR-Ab *p = 0.0121; g **p = 0.0023, and ***p = 0.000865; h *p = 0.029. i, j DIO mice were injected with saline or DA-GIP, and blood was collected 30 mins post-dose to measure plasma metabolites i corticosterone and j glycerol. n = 8 mice/group; one-way ANOVA with Sidak’s multiple comparisons test, i *p = 0.017 and **p = 0.0047, j *p = 0.043. Data are presented as mean values ± SEM. k, l DIO mice were pre-treated with vehicle or muGIPR-Ab 24-h before injection with saline or DA-GIP and plasma metabolites k corticosterone and l glycerol were measured over time. n = 10 mice/group; two-way repeated measures ANOVA with Sidak’s multiple comparisons test, k *p = 0.043, l *p = 0.029. Data are presented as mean values ± SEM. m, n cAMP concentration in mouse primary adipocytes (m) treated with GIP or (n) pre-treated with muGIPR-Ab overnight then treated with 10 nM mouse GIP. Data represents means ± SEM of three mice with three wells/treatment/mouse. o, p GIP-stimulated lipolysis in primary differentiated mouse pre-adipocytes o treated with mouse GIP compared to basal or p after pre-treatment with muGIPR-Ab overnight then treated with 10 nM mouse GIP. Data represent means ± SEM of 3–6 mice with three wells/treatment/mouse.