Journal: Cell Death & Disease

Article Title: Disrupted bone microenvironment and immune recovery following total body irradiation in a murine model

doi: 10.1038/s41419-025-08303-7

Figure Lengend Snippet: A Representative fluorescence images of the osteoclast phagocytosis assay in bone marrow–derived macrophages. BMM were stimulated with macrophage colony-stimulating factor (M-CSF), apoptotic thymocytes (APO), and receptor activator of nuclear factor kappa-B ligand (RANKL). Apoptotic thymocytes were labeled with CFDA (green), cytoplasm with CellMask Orange (orange), and nuclei with Hoechst 33342 (blue). Separate channels and merged overlays are shown. Cells with two nuclei were classified as pre-osteoclasts, and cells with three or more nuclei as osteoclasts. B Relative changes in the number, perimeter, and area of multinucleated TRAP+ cells after differentiation, with or without APO and transforming growth factor beta1 (TGF-β1) inhibitor. C Representative images of tartrate-resistant acid phosphatase (TRAP)-stained osteoclast cultures. D TRAP5b and E TGF-β protein expression in the supernatant of osteoclast cultures. F Analysis of relative gene expression of Nfatc1 (major transcription factor in osteoclastogenesis) , Acp5 (TRAP critical for osteoclastogenesis), and Rank (a receptor specific to osteoclasts and progenitors) gene. Statistical analysis involved a one-way ANOVA followed by Tukey’s multiple post hoc test. Data are presented as mean ± SD, and significance is indicated as * p < 0.05, ** p < 0.01, *** p < 0.001 ( n = 3–8).

Article Snippet: After 3 days of cultivation, TGF-β1 protein expression (Invitrogen 88-50690-22) and TRAP5b activity (Bone TRAP, Immunodiagnostic Systems) were measured in the culture supernatant from BMM differentiated osteoclasts.

Techniques: Fluorescence, Phagocytosis Assay, Derivative Assay, Labeling, Staining, Expressing, Gene Expression