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. 2020 Nov 13;11:5772. doi: 10.1038/s41467-020-19638-4

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 8 — a Cilia (AC3) staining in the hypothalamic ARH and hippocampal dentate gyrus of C57 neonates at P14 that received either saline or leptin antagonist SHLA injections from P4 to P13. The average lengths of 100 cilia and the ciliated cell percentage in each area per mouse are presented (n = 5 for hypothalamus, n = 4 for hippocampus). Scale bars: 20 μm. b Cilia staining in the hypothalamus (Hypo) and hippocampus (Hippo) of ob/+ mice, ob/ob mice, and ob/ob mice with leptin treatment (P10–P14) (n = 5). The average lengths of 100 cilia in each area per mouse are presented. Scale bars: 20 μm. c Upper panel: cilia (AC3) and POMC (β-END) costaining in the hypothalamic ARH in 7-week-old ob/+ mice, ob/ob mice, and ob/ob mice with adulthood leptin replacement (10 mg/kg/day for 7 days; n = 5). Lower panel: axonal projections of POMC neurons in the three groups (n = 4). Scale bars: 50 μm. d Ciliogenesis in the ARH and POMC axonal projection to the PVH in ob/ob neonates injected with saline, leptin alone, or leptin + shIFT88-GFP-AAV (n = 4 for AAV on-target group, n = 5 for the other 3 groups). The mice with off-target AAV injection were used as a control. Scale bars: 50 μm. e AC3 (cilia), functional leptin receptor (LepRb), GFP (AAV transfection), and DAPI costaining in the ARH of ob/ob neonates injected with saline, leptin, or leptin + shIFT88-GFP-AAV. The percentages of ARH cells and AAV-infected cells with periciliary leptin receptor accumulation were counted (n = 5 for the ob/ob-saline group and n = 4 for the other 2 groups). Scale bars: 10 μm. Data values are presented as mean ± SEM. Statistics were performed using two-sided Student’s t test (a) and one-sided one-way ANOVA (b–e) followed by post hoc LSD test. **p < 0.01, ***p < 0.001 between the indicated groups. ns not significant.