Figure 3. Decreased FoxO1 level in the hypothalamus of RIIβ KO mice.

(a) mRNA levels of FoxO1 in the hypothalamus of WT and RIIβ KO mice (WT, n=14; RIIβ KO, n=17, all male). (b) Representative immunoblots of hypothalamic FoxO1 and pAkt (Thr308) in fasted (24 h) WT and RIIβ KO mice after saline or leptin injection. Saline or leptin (1 mg kg⁻¹ body weight) was injected i.p. 30 min before mice were killed and the hypothalami were dissected and processed for western blot. (c) Quantification of protein levels of FoxO1 and pAkt by normalization to β-actin and total Akt, respectively. N values for WT:KO mice were FoxO1 saline (5:5), leptin (4:5) and pAkt/Akt saline (5:5). Values represent mean±s.e.m. *P<0.05, ***P<0.001 by Student's t-tests. Full blots are shown in Supplementary Fig. 3.