Fig. 6.
Characterization of hypothalamic signaling in LeprL536Hfs*6, wildtype, and Leprdb/db mice. A: Immunohistochemistry of pStat3 on brain slides. Wildtype (wt) +Leptin with pStat3 positive cells within VMH (ventromedial hypothalamus), DM (dorsomedial hypothalamic nucleus), and LH (lateral hypothalamic area), wt -Leptin, LeprL536Hfs*6 +/−Leptin, and Leprdb/db +/−Leptin without signal. Mice were fasted for 14–16 h prior to leptin injection (i.p. 5 μg/g body weight, +Leptin) or vehicle administration (−Leptin), 2 h later mice were sacrificed, brains were fixed by 4% PFA and removed, scale 200 μm. B: Induction of leptin signaling measured by western blot quantification of Jak2, Stat3, pAkt, Akt, Erk1/2, pErk1/2, and Socs3 in total hypothalamus protein. β-Actin served as loading control. Mice were fasted for 14–16 h prior to leptin injection (i.p. 5 μg/g body weight, +Leptin) or vehicle administration (−Leptin) and brains were removed 2 h later and hypothalamus was dissected out of the brain. Data shown as mean ± SEM, two-way ANOVA, n = 3/group, *P < 0.05.