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: Nature Communications

Article Title: Fusobacterium periodonticum promotes colorectal tumorigenesis via decanoic acid-driven neutrophil chemotaxis

doi: 10.1038/s41467-026-74591-y

Figure Lengend Snippet: A Flow cytometry analysis of neutrophil apoptosis after coculture with sodium decanoate (DeaNa), sodium palmitate (PaNa), and PBS. Quantitative graphs showing the proportion of late apoptotic/necrotic (Annexin V⁺/PI⁺) neutrophils ( B ) and early apoptotic (Annexin V⁺/PI⁻) neutrophils ( C ) under different treatments ( n = 3). D Schematic representation of neutrophil migration in vitro for 3 h at 37 °C. Created in BioRender. Yang, Q. (2026) https://BioRender.com/fm3t1j5 . Representative images of migrated neutrophils on the lower membrane stained with DAPI and photographed using the multifunctional enzyme reader. E Numbers of migrating neutrophils ( n = 3). F Volcano plot of differential gene expression in sodium decanoate-treated and PBS-treated neutrophils. G Heatmap showing the DEGs identified via RNA-seq in the presence of decanoic acid or PBS. H Quantitative reverse transcription PCR (qRT‒PCR) validation of gene expression changes in neutrophils treated with decanoic acid, palmitic acid and PBS ( n = 4). I Cell supernatant CXCL8 expression in neutrophils treated with decanoic acid, palmitic acid and PBS, as determined by ELISA ( n = 4). J Simplified schematic illustrating the coculture assay of neutrophils with Caco-2 cells, detailing the experimental setup. Created in BioRender. Yang, Q. (2026) https://BioRender.com/74ys0y4 . K qRT‒PCR validation of gene expression changes in Caco-2 cells after coculture with different fatty-acid-treated neutrophils ( n = 3). Data are shown as mean ± SEM, with representative results from at least three independent experiments. P values were determined by one-way ANOVA with Holm–Šidák test ( B , C ), one-way ANOVA with Dunnett’s test ( E , K ), one-way ANOVA with Tukey test ( H – I ). DeaNa, sodium decanoate; PaNa, sodium palmitate. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 and ns P > 0.05. Exact P values: B DeaNa vs PBS, P = 0.0465, PaNa vs PBS, P = 0.0002; C DeaNa vs PBS, P < 0.0001, PaNa vs PBS, P < 0.0001; E DeaNa vs PBS, P = 0.0342, PaNa vs PBS, P = 0.4726; H Cxcl2 , DeaNa vs PBS, P = 0.0015, PaNa vs PBS, P = 0.0001; Cxcl8 , DeaNa vs PBS, P = 0.0120, PaNa vs PBS, P = 0.0725; Cpt1a , DeaNa vs PBS, P = 0.0441, PaNa vs PBS, P < 0.0001; Osm , DeaNa vs PBS, P = 0.0078, PaNa vs PBS, P = 0.0019; Vegfa , DeaNa vs PBS, P = 0.0319, PaNa vs PBS, P = 0.0009. I DeaNa vs PBS, P = 0.0016, PaNa vs PBS, P = 0.0101. K CXCL8, DeaNa vs PBS, P < 0.0001, PaNa vs PBS, P = 0.9138; OSM, DeaNa vs PBS, P = 0.0865, PaNa vs PBS, P = 0.0031; VEGFA, DeaNa vs PBS, P = 0.0053, PaNa vs PBS, P < 0.0001; AREG, DeaNa vs PBS, P = 0.0451, PaNa vs PBS, P = 0.0391; PTGS2, DeaNa vs PBS, P = 0.0015, PaNa vs PBS, P = 0.0224.

Article Snippet: After treatment, cells were collected and stained using an Annexin V-FITC/Propidium Iodide (PI)-PE Apoptosis Detection Kit (MedChemExpress, USA).

Techniques: Flow Cytometry, Migration, In Vitro, Membrane, Staining, Gene Expression, RNA Sequencing, Reverse Transcription, Biomarker Discovery, Expressing, Enzyme-linked Immunosorbent Assay, Co-culture Assay