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. 2026 Jan 1;16(6):2780–2797. doi: 10.7150/thno.121905

Figure 2.

Figure 2

Osteogenic effects of osteocyte supernatants and characterization of osteocyte exosomes. (A) Lentivirus transfection of MLO-Y4 cells carrying GFP or Dll4 (GFP-osteocyte or Dll4-osteocyte). Scale bar = 100 μm. (B) qRT-PCR verified the expression changes of Dll4 gene in osteocytes. (C-D) ALP staining (C) and activity assay (D) of ST2 cells treated with 20% concentration of GFP-osteocyte or Dll4-osteocyte supernatants. Scale bar = 200 μm. (E) Morphology of exosomes isolated from osteocytes overexpressing Dll4 or GFP control (GFP-Exo or Dll4-Exo), observed under TEM. Scale bar = 200 nm. (F) NTA of GFP-Exo or Dll4-Exo. (G) Western blot of exosomal proteins (TSG101, HSP70, CD81, calnexin as a negative control) and Dll4 from both whole cell lysate and exosome lysate. (H) Representative immuno-EM images of GFP-Exo and Dll4-Exo co-stained with anti-Dll4 antibodies (conjugated with 10 nm gold particles, indicated by black arrows). Scale: 200 nm. (I) Levels of Dll4 expression in both GFP-Exo and Dll4-Exo as assayed by ELISA. *p < 0.05, **p < 0.01, ***p < 0.001 vs. GFP-Exo group.