Fig. 3: Overexpression of NT-3 in adipocytes promotes cold-induced thermogenesis and protects mice from diet-induced obesity.

a–c Nt-3 mRNA (a, WT n = 3, AdNT3-Tg n = 4, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), NT-3 protein levels measured by immunoblotting (b, WT n = 7, AdNT3-Tg n = 5, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test) and ELISA (c, n = 4/group, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test) in iWAT of WT and AdNT3-Tg mice. d–h Core body temperature (d, n = 5/group, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), fat pad mass (e, n = 4/group, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), Ucp1 and other thermogenic gene expression in iWAT (f, WT n = 4, AdNTF3-Tg n = 6, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), UCP1 protein levels in iWAT (g, n = 3, *indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), and representative UCP1 immunostaining images showing UCP1-positive beige adipocytes in iWAT (h, from three replicate animals/group, Scale bar for the ×40 images = 75 µm, and for the ×20 images = 150 µm) of AdNT3-Tg mice and their littermates during cold exposure. i–l Body weight (i, WT n = 8, AdNT3-Tg n = 10, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), fat pad mass (j, n = 7/group, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), Oxygen consumption (k, WT n = 5, AdNT3-Tg n = 6, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test) and heat production (l, WT n = 5, AdNT3-Tg n = 6, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test) in AdNT3-Tg mice and their littermate controls fed with HFD starting 6 weeks of age. m–o) Ucp1 and other thermogenic gene expression (m, WT n = 7, AdNTF3-Tg n = 8, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), UCP1 and TH protein levels (n, n = 8/group, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), and representative UCP1 immunostaining in iBAT (o, from three replicate animals/group, scale bar = 75 µm) of AdNT3-Tg mice and their littermate controls fed HFD. p, q Ucp1 and other thermogenic gene expression (p, WT n = 6, AdNTF3-Tg n = 7, * indicates statistical significance between WT and AdNT3-Tg with unpaired two-tailed t test), and H&E staining (q, from three replicate animals/group, scale bar = 150 µm) in iWAT of AdNT3-Tg mice and their littermate controls fed a HFD. All data are expressed as mean ± SEM.