Journal: Bioactive Materials

Article Title: Screening of a quinonoid compounds library identifies decylubiquinone as an antioxidant and anti-apoptotic agent against glucocorticoid-induced osteoporosis via CD39/CD73/adenosine axis

doi: 10.1016/j.bioactmat.2026.03.062

Figure Lengend Snippet: Roles of the CD39/CD73 axis on DUB's multidirectional protection effects in Dex-treated primary BMSCs. ( A ) ELISA for ROS clearance-related enzyme T-SOD and ROS damage biomarkers 8-OHdG, AOPP, and MDA in primary BMSCs of different groups. ( B ) Western blot and quantification for the expression of ROS clearance-related proteins in primary BMSCs of different groups. ( C ) Representative images and quantitative analysis of immunofluorescence staining for MitoSox (red) in primary BMSCs of different groups, and nuclei were stained with Hoechst (blue). ( D ) Western blot and quantification for the expression of apoptosis-related proteins in primary BMSCs of different groups. ( E ) Cellular apoptosis detection in primary BMSCs of different groups by Annexin V-FITC and PI dual-staining assessment via flow cytometry. The proportion of cells in each quadrant was indicated in the plot. ( F ) Tunel (red) staining and quantification of apoptotic cells in primary BMSCs of different groups, and nuclei were stained with DAPI (blue). ( G ) Representative images and quantitative analysis of Alizarin Red S staining for mineralization in primary BMSCs of different groups under osteogenic conditions. ( H ) Western blot and quantification for the expression of osteogenesis-related proteins in primary BMSCs of different groups. n = 4 independent repeats by using different biological samples in each group for in vitro experiments. Data were means ± s.e.m. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by one-way ANOVA. Scale bars: 50 μm (C), 25 μm (F), and 200 μm (G).

Article Snippet: The proportion of early and late apoptotic primary BMSCs under different treatment conditions was determined using an Annexin V-FITC/PI Apoptosis Detection Kit (E-CK-A211, Elabscience, Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence, Staining, Flow Cytometry, TUNEL Assay, In Vitro

Journal: Bioactive Materials

Article Title: Screening of a quinonoid compounds library identifies decylubiquinone as an antioxidant and anti-apoptotic agent against glucocorticoid-induced osteoporosis via CD39/CD73/adenosine axis

doi: 10.1016/j.bioactmat.2026.03.062

Figure Lengend Snippet: The effect of ADO supplements on primary BMSCs. ( A ) MTT assay for the proliferation of BMSCs treated with different doses of ADO for 2 days and 10 days under osteogenic induction conditions with or without 10 μM Dex. ( B-C ) Representative images and quantitative analysis of mineralized nodule areas by Alizarin Red S staining in primary BMSCs treated with gradient doses of ADO under osteogenic induction with or without 10 μM Dex. ( D ) Western blot and quantification for the expression of osteogenesis-related proteins in primary BMSCs of different groups. ( E ) ELISA for ROS clearance-related enzyme T-SOD and ROS damage biomarkers 8-OHdG, AOPP, and MDA in primary BMSCs of different groups. ( F ) Western blot and quantification for the expression of ROS clearance-related proteins in primary BMSCs of different groups. ( G ) Representative images and quantitative analysis of immunofluorescence staining for MitoSox (red) in primary BMSCs of different groups, and nuclei were stained with Hoechst (blue). ( H ) Western blot and quantification for the expression of apoptosis-related proteins in primary BMSCs of different groups. ( I ) Cellular apoptosis detection in primary BMSCs of different groups by Annexin V-FITC and PI dual-staining assessment via flow cytometry. The proportion of cells in each quadrant was indicated in the plot. ( J ) Tunel (red) staining and quantification of apoptotic cells in primary BMSCs of different groups, and nuclei were stained with DAPI (blue). n = 4 independent repeats by using different biological samples in each group for in vitro experiments. Data were means ± s.e.m. ∗∗ p < 0.01, ∗∗∗ p < 0.001 by one-way ANOVA. Scale bars: 200 μm (B), 50 μm (G), and 25 μm (J).

Article Snippet: The proportion of early and late apoptotic primary BMSCs under different treatment conditions was determined using an Annexin V-FITC/PI Apoptosis Detection Kit (E-CK-A211, Elabscience, Wuhan, China).

Techniques: MTT Assay, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Flow Cytometry, TUNEL Assay, In Vitro

Journal: Bioactive Materials

Article Title: Screening of a quinonoid compounds library identifies decylubiquinone as an antioxidant and anti-apoptotic agent against glucocorticoid-induced osteoporosis via CD39/CD73/adenosine axis

doi: 10.1016/j.bioactmat.2026.03.062

Figure Lengend Snippet: Summary of the study. The schematic diagram illustrates that DUB alleviates GIOP by suppressing oxidative stress and apoptosis via the CD39/CD73/ADO axis and promotes osteogenesis via ADO/A 2b R-mediated activation of the PKA/CREB pathway. The schematic diagram was created by using BioRender.com.

Article Snippet: The proportion of early and late apoptotic primary BMSCs under different treatment conditions was determined using an Annexin V-FITC/PI Apoptosis Detection Kit (E-CK-A211, Elabscience, Wuhan, China).

Techniques: Activation Assay

Journal: Molecular Therapy Oncology

Article Title: The miR-16-1-3p suppresses proliferation and invasiveness via the MDM2-p53 axis in TGF-β1 signaling in osteosarcoma

doi: 10.1016/j.omton.2026.201249

Figure Lengend Snippet: miR-16-1-3p counteracts TGF-β1-mediated effects on metastasis and chemosensitivity of OS cells (A) Schematic representation of the chick embryo chorioallantoic membrane (CAM) assay. Stably Katushka2S-expressing U2OS cells were implanted onto the CAM to evaluate tumor growth and metastasis. Eggs were imaged at the indicated time points, and tumors were quantified at the experimental endpoint using a fluorescence imaging system. (B) Representative fluorescent and bright-field macroscopic images of tumor nodules formed on the CAM (left), with quantification of tumor volume shown on the right. (C) Representative fluorescent images of metastatic dissemination formed by Katushka2S-labeled U2OS cells in chick embryos under the indicated treatment conditions at the experimental endpoint. (D) Immunofluorescence analysis of Ki67 expression in cryosections of CAM tumor nodules. Representative images of Ki67 staining (magenta), DAPI-positive nuclei (blue), and merged channels are shown (left). Quantification of Ki67-positive cells as a percentage of total cells is shown on the right. Scale bars, 150 μm. (E) SRB assay of dose-dependent proliferation responses of U2OS cells to cisplatin under the indicated treatment conditions. Dose-response curves are shown in the upper panel, and calculated IC50 values are listed below. (F) Flow cytometric analysis of apoptosis using Annexin V-FITC/PI staining following cisplatin treatment. Representative dot plots are shown (top), with quantification of apoptotic cells shown on the bottom. (G) Kaplan-Meier survival analysis of TARGET-OS patients stratified according to combined TGFB1 and MDM2 expression levels. Patients with high TGFB1 and low MDM2 expression showed the most favorable survival outcome. Data are presented as mean ± SD. p values in (B, D, and F) were determined using one-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns, not significant.

Article Snippet: Apoptosis was evaluated using an Annexin V-FITC/PI Apoptosis Detection Kit (Servicebio, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Membrane, Chick Chorioallantoic Membrane Assay, Stable Transfection, Expressing, Fluorescence, Imaging, Labeling, Immunofluorescence, Staining, Sulforhodamine B Assay