Extended Data Fig. 7: Pre-treatment with mGIPR-Ab and mGIPR-Ab/P3 blocks DA-GIP- induced c-FOS in the mouse brain.
GIPR and GLP-1R expressions were visualized in the area postrema by in situ hybridization. (a) mGIPR (pink) and mGLP1R (teal) were localized on different cells, but within proximity of each other, in the area postrema. Relative expression levels of mGIPR and mGLP1R were scored on a scale of 0-4 based on ACD scoring criteria, where level 2 indicates ‘4-9 dots/cell, no or very few dot clusters’. The effect of mGIPR-Ab and mGIPR-Ab/P3 on DA-GIP induced c-Fos in the area postrema was analyzed with fluorescent immunohistochemistry. (b, e) DA-GIP significantly increases c-Fos expression in the area postrema following IgG1 pre-treatment. (c, e) DA-GIP expression does not increase c-Fos expression in the area postrema with mGIPR-Ab pre-treatment. (d, e) Moreover, DA-GIP does not increase c-Fos expression in the area postrema following mGIPR-Ab/P3 pre-treatment, however, mGIPR-Ab/P3 alone induces high levels of c-Fos expression in the area postrema. (A) n = 2, (B-E) IgG1/A52Su (n=5), IgG1/DA-GIP (n=4), mGIPR-Ab/A52Su (n=6), mGIPR-Ab/DA-GIP (n=6), mGIPR-Ab/P3/A52Su (n=6), mGIPR-Ab/DA-GIP (n=6). One-way ANOVA with Sidak’s test for multiple comparisons; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 comparison indicated by bracket. Data represent mean ± SEM. Scale bar 100 µm.
