Fig. 4

MANF overexpression in the hypothalamus led to increased body weight and food intake. a Schematic representation of AAV-MANF construct. b A single western blotting analysis of viral MANF expression in different brain regions of 2-month old wild type (WT) mice injected with AAV-MANF into the hypothalamus. c Immunofluorescence images of 2-month-old WT mice injected with AAV-GFP or AAV-MANF into the hypothalamus. Green fluorescence was used to indicate the expression of GFP in different regions of the hypothalamus, including arcuate nucleus (ARC), dorsomedial hypothalamus (DMH) and lateral hypothalamus (LH). Red fluorescence was used to indicate the expression of MANF, which was stained by an HA antibody. Higher magnification (63×) images revealed cytoplasmic localization of MANF. Nuclear staining is shown in the merged image. Staining of hypothalamus from uninjected mice was used as negative controls (Scale bar: left 50 μm, right 10 μm). d A single image of WT mice 2 weeks after AAV-GFP or AAV-MANF injection. e Body weights of male and female 2-month-old WT mice were measured weekly before and after AAV-GFP or AAV-MANF injection (*P < 0.05, **P < 0.01, n = 5, two-way ANOVA with Bonferroni post-tests, Male, F = 153.3, P < 0.0001; Female, F = 74.98, P < 0.0001). f Food intake of male and female 2-month-old WT mice were measured weekly before and after AAV-GFP or AAV-MANF injection (**P < 0.01, ***P < 0.001, n = 5, two-way ANOVA with Bonferroni post-tests, Male, F = 204.8, P < 0.0001; Female, F = 524.2, P < 0.0001). Data are represented as mean ± SEM