Extended Data Fig. 2: NachBac expression in ArcGABA+ neurons increases neuron activity levels and promotes massive obesity.
From: Profound and redundant functions of arcuate neurons in obesity development
a, A represent expression of pattern of NachBac vector in the Arc from rostral to caudal sections. b, Expression GFP (green column) and c-Fos (red column) and their colocalization (merged column) with magnified pictures showing the boxed area in the merged pictures from Vgat-Cre mice injected with Cre-dependent GFP (top two rows) or NachBac (bottom two rows) when fed ad libitum chow (the first and third rows) or fasted (the second and forth row). It is notable that the vast majority of NachBac-expressing neurons exhibited c-Fos regardless of fed or fasted. This is one of 3 animals/group shown in Fig. 1f. c-g, Associated data from animals presented in Fig. 1g–k, n = 6 animals/group. Comparison in lean mass between control and mice with NachBac expression in ArcGABA-NachBac mice 11 weeks after viral delivery (n = 6 each/group, t = 1.055, df=10, p = 0.3163, 2-sided unpaired Student’s t test). d, Comparison in body weight when mice were placed CLAMS chambers for measurements during the 1st week after viral delivery (n = 6/group, t = 0.7030 df=10, p = 0.4981, 2-sided unpaired Student’s t test). (e–g) Real time measurements of food intake (e), O2 consumption (f) and locomotion (g) over the 3 day measurement period. h, Comparison in leptin levels between groups 11 weeks after viral delivery (n = 6/group, t = 14.88 df=10, **p < 0.0001, 2-sided unpaired Student’s t test). i–m, Female Vgat-Cre were injected with AAV-Flex-EGFP-P2A-mNachBac or control GFP vectors to bilateral Arc at 7–8 weeks of age. Body weight (i, n = 6 each/group, df=11, F(11, 84)=41.03, **p < 0.0001, two-way ANOVA tests) and body weight gain (j, n = 6 each/group, df=11, F(11, 84)=38.60, **p < 0.0001, two-way ANOVA tests) were followed weekly for 11 weeks. k–m, Comparison in food intake measured at the first 2 weeks after viral delivery (k, t = 6.700 df=7, **p = 0.0003, 2-sided unpaired Student’s t test), lean mass (l, t = 0.9398 df=7, p = 0.3786, 2-sided unpaired Student’s t test) and fat mass (m, t = 19.46 df=7, **p < 0.0001, 2-sided unpaired Student’s t tests) measured at the end of the 11th week after viral delivery. 3 V: the third ventricle. All data presented as mean + /-sem.
