Fig. 7

Colocalization and effects of direct NPFFR2 signalling on NPY neurons. a Representative image of GFP expression in the Arc of a NPY-TRAP mouse brain. Scale bar = 100 µm. b, c Quantification of the expression of Npy and Npffr2 mRNA in the input and immunoprecipitated (IP) RNA isolated from the Arc of NPY-TRAP (n = 10), Ins-TRAP (n = 3) and WT-TRAP (n = 3) mice. One-way ANOVA was used to determine difference between groups. ^∗∗p < 0.05 as indicated by bar. N.E, gene is not expressed. d Double chromogenic ISH for Npy (red) and Npffr2 (blue) expression in the Arc. Scale bar = 100 µm in the image on the left, 10 µm in images (i), (ii) and (iii). Representative image from three independent experiments. e Representative images of IHC of brain sections for pCREB (magenta) in NPY-GFP mice that received i.c.v. either saline, NPFF (150 µM), Forksolin (2.5 mM) or NPFF (150 µM) + Forksoline (2.5 mM). Brains were collected 30 min after i.c.v. injection. Scale bar = 100 µm. f Quantification of pCREB-ir-positive neurons colocalized with Arc GFP-labelled NPY neurons expressed as percentage of Arc NPY neurons. Forskolin n = 5, Forskolin + NPFF n = 4. Data are mean ± SEM. One-way ANOVA was used to determine difference between groups. ^∗∗p < 0.05 as indicated by bars