Fig. 5

NPFFR2 signalling regulates the Arc NPY – PVN-TH pathway. a pSTAT3-ir cells in the Arc of male WT and Npffr2^−/− mice fed on either chow or 6 weeks of HFD. Brains were collected at 30 min after an i.p. injection of saline or leptin (2 µg/g body weight). Another cohort of these mice received intracerebroventricular (i.c.v.) injection of saline or leptin (1.25 µg) at 30 min prior to perfusion and brain collection. n = 3–4 brains per group. Scale bar = 100 µm. b Representative images of IHC on pSTAT3 used for pSTAT3-ir cell quantification after i.p. injection of either saline or leptin shown in a. c, d qPCR analysis on the mRNA expression of Npffr2 in hypothalamus (HT), brown adipose tissue (BAT), subcutaneous white adipose tissue (WAT), hypothalamic expression on chow and 9 weeks of HFD. n = 3–4 per group. e Image of chromogenic ISH for Npffr2 expression at the Arc. Scale bar = 100 µm in the image on the left, 50 µm in images (i) and (ii). Representative images from 3 independent experiments. f, g, h The expression of Npy and Pomc by ISH at Arc, and Th at PVN in WT and Npffr2^−/− mice fed on chow or 9 weeks of HFD. n = 5–7 per group. i Representative images of ISH on Npy expression at the Arc quantified in f. Scale bar = 100 µm. j Representative images of ISH on Th expression at the PVN quantified in h. Scale bar = 100 µm. Data are mean ± SEM. One-way ANOVA was used to determine the treatment (a), tissue type (c) diet and genotype (d, b, g, h) effects among groups. ^∗∗p < 0.05 as indicated by bar. Arc, arcuate nucleus. PVN, paraventricular nucleus of the hypothalamus