Fig. 8: ONECUT3 loss-of-function in the PeVN of the mouse hypothalamus.

a siRNA-mediated Onecut3 (OC3) knock-down reduced ONECUT3 protein levels in the nuclei (open arrowheads) of Onecut3-mCherry⁺ neurons in the LH of neonatal mice. Mice were unilaterally injected in the LH on P4 and sampled on P10. ONECUT3 protein content was visualized by immunohistochemistry. a₁ ONECUT3 (OC3) signal intensity was compared between the non-injected (ctrl) vs siRNA-injected hemispheres of mice (n = 4/group; p = 0.0431). Data from each mouse were interconnected. a₂ The number of ONECUT3⁺ neurons that populated the LH did not differ as a result of siRNA injection (ns non-significant; n = 4 subjects/group). b–b₂ In situ hybridization revealed the downregulation of Nav2 mRNA after siRNA treatment (dotted lines denote the individual cells measured, n = 3 subjects/group, 15 cells; p = 0.0433 (b₁) and 0.0363 (b₂). c, c₁ Knock-down of Onecut3 reduced neurite density, as measured by the area coverage of mCherry⁺ fibers in the LH, noting that Onecut3⁺ neurons were multipolar. The area occupancy of mCherry⁺ neurites, likely local dendrites, in the LH of Onecut3-mCherry mice, was compared between the non-injected (ctrl) vs siRNA-injected hemispheres (n = 4 mice; p < 0.05). Data from each mouse were interconnected. p = 0.0497, n.s. non-significant, two-tailed Student’s t-test. Scale bars = 250 µm (a), 20 µm (b), and 100 µm (c). Source data are provided as a Source Data file.