Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Jul 10;632(8024):357–365. doi: 10.1038/s41586-024-07634-3

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 5 — a, Left, schematic of experiment to induce loss-of-function of CCN3 (Ccn3 knockdown) in the ARC in mice. Right, Ccn3-positive neurons in female Esr1^Nkx2.1-cre ARC versus %BV/TV after Ccn3 siRNA injections. b, Ccn3 expression in control, unilateral and bilateral hit with corresponding µCT scans of distal femurs. Scale bar, 500 µm (N = 6, 4). c, Left, schematic of experiment to induce gain-of-function CCN3 in the liver. Right, ectopic mCCN3 expression in Esr1^fl/fl female hepatocytes following retro-orbital injection of AAVdj-CAG-CCN3 (AAVdj-CCN3) or control (AAVdj-Ctrl) vectors. Inset shows double nuclei. Scale bar, 100 µm. d, mCCN3 immunoblot of heparin–agarose-purified plasma (left) and liver extracts (right, 10 µg total protein) from mice 5 weeks after injection with AAVdj-Ctrl (–) or AAVdj-mCCN3 (+). Recombinant mCCN3 (rCCN3) shown in far left lane. e, %BV/TV (left) of femurs and L5 and mechanical strength (right) of femurs from 3-4-month-old Esr1^fl/fl female mice 5 weeks after injection (N = 7, 8 femurs and N = 4, 3 L5). f, %BV/TV (left) of femurs and L5 and mechanical strength (right) of femurs from 3–4-month-old Esr1^fl/fl males 5-weeks after injection (N = 6, 7 femurs and N = 5, 7 L5). g, %BV/TV of femurs and L5 in 5-month-old OVX Esr1^fl/fl females 9 weeks after injection (N = 8, 8 femurs and N = 8, 8 L5). h, %BV/TV of 20–23-month-old Esr1^fl/fl female femurs 9 weeks after injection (N = 5, 5). i, Bone formation rate and bone surface (BFR/BS), and number of osteoblasts per bone surface (No. Ob/BS) determined by histomorphometry (N = 7, 8). Scale bar, 50 µm. j, Number of osteoclasts per bone surface (No. Oc/BS) and lacunar density per bone area as determined by static histomorphometry (N = 4, 4). Representative images of TRAP (top) and silver nitrate (AgNO3, bottom) staining for femoral osteoclasts and lacunae, respectively. Scale bar, 50 µm. Simple linear regression for a. Unpaired Student’s t-test, one-tailed or two-tailed for e–g, i and j as indicated in Supplementary Table 3. Mann–Whitney test, one-tailed for h. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Error bars ± s.e.m. Graphic in c was adapted from BioRender (https://www.biorender.com).

Source Data