annexin v fitc pi apoptosis detection kit  (Beyotime)

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: Bioactive Materials

Article Title: Three-dimensional printed PCL/nHA scaffolds promote soft tissue functional fibrosis to repair chest wall defect via Piezo1/Ca 2+ signal during respiratory motion

doi: 10.1016/j.bioactmat.2026.03.037

Figure Lengend Snippet: 10 wt% PCL/nHA enhances cell proliferation and reduces apoptosis via calcium ion regulation. (A) Morphological characteristics and viability of L929 cells cultured on PCL and PCL/nHA samples. (B) Statistical evaluation of L929 cells viability. (C) Morphological characteristics and viability of HSFs cultured on PCL and PCL/nHA samples. (D) Statistical evaluation of HSFs viability. (E) Time-dependent proliferation of L929 cells on PCL and PCL/nHA substrates over 1, 2, and 3 days. (F) Time-dependent proliferation of HSFs on PCL and PCL/nHA substrates over 1, 2, and 3 days. (G) Influence of varying nHA concentrations in PCL on L929 cells apoptosis. (H) Quantitative apoptosis assessment across experimental groups based on data from G. (I) Apoptosis of L929 cells cultured in media with varying calcium ion concentrations. (J) Statistical analysis of the apoptosis data presented in I. (K) The morphological characteristics and viability of L929 cells cultured in the CaCl 2 group, the nHA group (with calcium ion concentration equivalent to that in the CaCl 2 group), and the control group. All data are presented as the Mean ± SD (n ≥ 3). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001. ns, no significant difference.

Article Snippet: CCK-8 assay kit, Fluo-4AM, Cell Apoptosis Detection Kit, Mitochondrial membrane potential assay kit and ROS Assay Kit were purchased from Beyotime Biotechnology (China).

Techniques: Cell Culture, Concentration Assay, Control

Journal: Bioactive Materials

Article Title: Three-dimensional printed PCL/nHA scaffolds promote soft tissue functional fibrosis to repair chest wall defect via Piezo1/Ca 2+ signal during respiratory motion

doi: 10.1016/j.bioactmat.2026.03.037

Figure Lengend Snippet: Piezo1-mediated Ca 2+ signaling in PCL/nHA-Driven cell behaviors. (A) Fluorescence imaging of intracellular calcium ion influx in L929 cells on PCL and PCL/nHA surfaces. (B) Fluorescence quantification of calcium ion influx in A. (C) Fluorescence imaging of intracellular calcium ion influx in HSFs on PCL and PCL/nHA surfaces. (D) Fluorescence quantification of calcium ion influx in C. (E) Fluorescence imaging assessment of intracellular ROS levels in L929 cells on PCL and PCL/nHA samples. (F) Effects of PCL samples with different nHA contents on the mitochondrial membrane potential of L929 cells. (G) Assessment of the expression levels of mechanosensing- and proliferation-related proteins in L929 cells cultured on PCL and PCL/nHA surfaces. (H) Assessment of the expression levels of mechanosensing- and proliferation-related proteins in HSFs cultured on PCL and PCL/nHA surfaces. (I) Assessment of the expression levels of apoptosis-related proteins in L929 cells cultured on PCL and PCL/nHA surfaces. (J) Schematic illustration depicting the mechanisms underlying L929 cell proliferation differences. All data are presented as the Mean ± SD (n ≥ 3). ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001. ns, no significant difference.

Article Snippet: CCK-8 assay kit, Fluo-4AM, Cell Apoptosis Detection Kit, Mitochondrial membrane potential assay kit and ROS Assay Kit were purchased from Beyotime Biotechnology (China).

Techniques: Fluorescence, Imaging, Membrane, Expressing, Cell Culture