Figure 6. Obesity in Nkx2-1f/f/Sf1Cre females is associated with fewer NKX2-1+, ERα+ and Tac1+ neurons in the VMHVL.
(A) NKX2-1 (red) and ERα (green) immunoreactivity in the VMH and ARC of P10 Nkx2-1f/f/Sf1Cre and Nkx2-1f/f mice. Images show coronal brain sections, VMH, VMHVL, and ARC bordered by dashed lines, scale bars = 100 µm, 3V = third ventricle.
(B) Quantification of ERα+ nuclei in the VMHVL or ARC from P10 Nkx2-1f/f/Sf1Cre (green) or Nkx2-1f/f female mice (black) (n = 2 mice/group, 3 sections/brain). Bottom panel shows double-labeling of NKX2-1 (green) and ERα (red) in the VMHVL of P110 control females. ERα-negative/NKX2-1-positive (arrows) and ERα-positive/NKX2-1-negative nucleus (asterisk).
(C) Quantification of cells derived from the Sf1 lineage determined for the ventrolateral (VMHVL), central (VMHc), and dorsomedial (VMHdm) subregions of Nkx2-1f/f/Ai14f/+/Sf1Cre mutant (red) and Nkx2-1f/+/Ai14f/+/Sf1Cre control (black) P10 female mice (n = 5 animals/group, 3 sections/brain).
(D) Relative expression by qPCR of Nkx2-1, Nmu, Tac1, Esr1, and Gal from microdissected VMH of P10 Nkx2-1f/f/Sf1Cre (grey) or Nkx2-1f/f (black) females (n = 3–6/group).
(E) Double-labeling of NKX2-1 immunoreactivity (green, nuclear) and Tac1 transcripts (red, cytoplasmic) analyzed by confocal microscopy in the VMHVL of P10 wild-type female mice.
(F) Patterns of Esr1 and Tac1 transcripts by ISH in Nkx2-1f/f/Sf1Cre or Nkx2-1f/f P10 females.
Pairwise comparisons performed with Holm-Sidak multiple comparison tests following two-way ANOVA, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. See also Figure S6.