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. 2023 Apr 17;133(8):e162519. doi: 10.1172/JCI162519

Figure 4. Effects of local Sn osteocyte clearance on empty lacunae, osteocyte apoptosis, and osteocyte LCN quality.

Figure 4

(A) Study design for the local clearance of Sn osteocytes in old (20 months) DMP1-Cre+/– p16-LOX-ATTAC mouse cohorts, males and females combined, randomized to Veh (gray) or AP (teal) treatment for 4 months. (B–D) Study endpoint (24 months) quantification of (B) total osteocytes per bone area and (C) percentage of osteocyte empty lacunae in mice treated with Veh (n = 8: n = 4 females, n = 4 males) versus AP (n = 8: n = 4 females, n = 4 males), and (D) representative images of osteocyte empty lacunae (arrowheads). Scale bar: 25 μm. (E–G) Quantification of osteocyte apoptosis, including (E) total numbers of osteocytes analyzed and (F) percentage of osteocyte apoptosis in mice treated with Veh (n = 8: n = 4 females, n = 4 males) versus AP (n = 8: n = 4 females, n = 4 males), and (G) representative images of DAPI-stained, TUNEL+, and merged apoptotic osteocytes. Scale bars: 25 μm. (H) Quantification of osteocyte LCN score for mice treated with Veh (n = 9: n = 5 females, n = 4 males) versus AP (n = 5: n = 3 females, n = 2 males), and (I) representative images of DAPI-stained, phalloidin-stained, and merged osteocyte LCNs. Scale bars: 25 μm. Data represent the mean ± SEM. NS, P > 0.05; *P < 0.05, by independent samples Student’s t test or Wilcoxon rank-sum test, as appropriate.