tunel assay kit  (Servicebio Inc)

Journal: Poultry Science

Article Title: Paeonol alleviates granulosa cell senescence in laying chickens via the PI3K/Akt/mTOR signaling pathway

doi: 10.1016/j.psj.2026.106750

Figure Lengend Snippet: Paeonol alleviates D-gal-induced apoptosis in GCs. A-E: qRT-PCR detection of BCL2 , Caspase-3, PCNA, CDK2 , and CCND1 . F-K: Western blot detection of Bax, Bcl-2, PCNA, Caspase-3, and CDK2 proteins. L-M: Annexin V/PI staining and flow cytometry analysis for detection of apoptosis; Q2: Annexin V⁺ / PI⁺, late apoptotic cells; Q3: Annexin V⁺ / PI⁻, early apoptotic cells. Data are presented as mean ± SEM ( n ≥ 3). Different lowercase letters indicate significant differences ( P < 0.05).

Article Snippet: Deparaffinized sections were processed using a BrightGreen Apoptosis Detection Kit (A112-03, Vazyme, Nanjing, China) according to the manufacturer's protocol.

Techniques: Quantitative RT-PCR, Western Blot, Staining, Flow Cytometry

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: Additionally, apoptosis was independently assessed by using a One-step TUNEL In Situ Apoptosis Kit (E-CK-A322, Elabscience, Wuhan, China) according to the manufacturer's instructions.

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: Additionally, apoptosis was independently assessed by using a One-step TUNEL In Situ Apoptosis Kit (E-CK-A322, Elabscience, Wuhan, China) according to the manufacturer's instructions.

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: Additionally, apoptosis was independently assessed by using a One-step TUNEL In Situ Apoptosis Kit (E-CK-A322, Elabscience, Wuhan, China) according to the manufacturer's instructions.

Techniques: Activation Assay

Journal: Materials Today Bio

Article Title: A dual-responsive CO-releasing nanogel ameliorates retinal ischemia–reperfusion injury by restoring mitochondrial homeostasis and attenuating cGAS-STING pathway activation

doi: 10.1016/j.mtbio.2026.102974

Figure Lengend Snippet: COPN mitigates oxidative stress, neuroinflammation, and apoptosis in OHT-induced RIRI. a–f , Quantitative analyses of apoptotic markers (TuJ-1, Brn3A, Caspase-3, Bax, Cytochrome-c, and Bcl-2) in retinas subjected to various treatments, which revealed a significant reduction in COPN-induced apoptosis. g , TUNEL assay demonstrating reduced apoptosis in COPN-treated retinas (red fluorescence indicates apoptotic cells, and nuclei are stained with DAPI, blue). h–o , ELISA quantification of proinflammatory cytokines (TNF-α, IL-1β, IFN-γ, IL-18, and MCP-1) and oxidative stress markers (CAT, SOD, MDA, and MCP-1), demonstrating the broad anti-inflammatory and antioxidative effects of COPN. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed via the TUNEL BrightRed Apoptosis Detection Kit (Vazyme, Cat# A113-01) according to the manufacturer's instructions.

Techniques: TUNEL Assay, Fluorescence, Staining, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: A dual-responsive CO-releasing nanogel ameliorates retinal ischemia–reperfusion injury by restoring mitochondrial homeostasis and attenuating cGAS-STING pathway activation

doi: 10.1016/j.mtbio.2026.102974

Figure Lengend Snippet: COPN mitigates retinal ischemia‒reperfusion injury by suppressing the cGAS–STING–TBK1 signaling axis. a , Heatmap illustrating the differential expression of key genes involved in mitochondrial function, apoptosis, and inflammatory responses between OHT-treated and control retinas. b , Gene Ontology (GO) enrichment analysis highlighting significantly enriched biological processes and molecular functions, including the mitochondrial stress response, cGAS-STING complex activation, and neuroinflammation. c , Volcano plot depicting significantly differentially expressed genes (DEGs) between the OHT and control groups, with upregulated genes in red and downregulated genes in blue. d , KEGG pathway enrichment analysis revealed significant activation of oxidative phosphorylation disruption, apoptosis, inflammation, and notably the cGAS‒STING signaling pathway in retinal ischemia. e , RT‒qPCR quantification of key components of the cGAS‒STING pathway (cGAS, STING, TBK1, IRF3, and caspase-3) in the retinas of mice after different treatments. f–g , Western blot analyses of cGAS-STING pathway-related proteins (cGAS, STING, p-STING, TBK1, p-TBK1, IRF3, p-IRF3, and cleaved caspase-3) under (f) OGD/R stress in R28 cells and (g) retinal tissues from OHT mice. ∗∗∗∗ P < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed via the TUNEL BrightRed Apoptosis Detection Kit (Vazyme, Cat# A113-01) according to the manufacturer's instructions.

Techniques: Quantitative Proteomics, Control, Activation Assay, Phospho-proteomics, Disruption, Western Blot

Journal: Bioactive Materials

Article Title: Ameliorating post-infarction myocardial fibrosis and cardiac function via ROS-responsive hydrogel-mediated IL-11 antibody delivery

doi: 10.1016/j.bioactmat.2026.01.041

Figure Lengend Snippet: The expression of interleukin-11 (IL-11) is increased in myocardial fibrosis in vitro and in vivo. A. Western blot analysis of IL-11 and vimentin expression in angiotensin II (AngII, 100 nM) or TGF-β (10 ng/mL)-treated cardiac fibroblasts. B. Quantification of (A). C. TUNEL staining and (D) its quantitative analysis in treated cells. E. CCK-8 viability assays (n = 3). F. Circulating IL-11 levels in mouse blood measured by ELISA (n = 3). ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Sections were fixed in 4 % PFA for 30 min, permeabilized, and blocked with DAKO solution (0.1 % saponin) for 1 h. Apoptosis was detected using the One Step TUNEL Apoptosis Kit (Beyotime, C1089) per manufacturer's instructions.

Techniques: Expressing, In Vitro, In Vivo, Western Blot, TUNEL Assay, Staining, CCK-8 Assay, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Ameliorating post-infarction myocardial fibrosis and cardiac function via ROS-responsive hydrogel-mediated IL-11 antibody delivery

doi: 10.1016/j.bioactmat.2026.01.041

Figure Lengend Snippet: P-T@MAB injection promotes angiomyogenesis. Immunostaining images show TUNEL at 3 days post-injection (A), Ki67 (C), vWF (E), and CD31 (F) expression in heart sections at 4 weeks post-injection. Quantitative analysis of TUNEL (B), Ki67 (D), vWF (G), and CD31 (G) is presented (n = 5). Scale bars: 100 μm. n.s., not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Sections were fixed in 4 % PFA for 30 min, permeabilized, and blocked with DAKO solution (0.1 % saponin) for 1 h. Apoptosis was detected using the One Step TUNEL Apoptosis Kit (Beyotime, C1089) per manufacturer's instructions.

Techniques: Injection, Immunostaining, TUNEL Assay, Expressing

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Schematic diagram illustrates the mechanism of Mn-NC composite hydrogels for TMJ-OA treatment. Composite hydrogels can effectively protect chondrocytes from ROS damage, alleviate inflammatory reactions within the joint microenvironment, inhibit inflammation-induced chondrocyte apoptosis and ECM degradation, and simultaneously induce the regeneration and repair of subchondral bones via the enzyme-like activity of Mn-NC SAzymes and the release of Mn ions.

Article Snippet: TUNEL staining was performed using a TUNEL Apoptosis Assay Kit (red fluorescence; Beyotime Biotechnology Company, China).

Techniques: Activity Assay

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The Mn-NC composite hydrogels inhibit chondrocyte apoptosis and ECM degradation in rats after IL-1β stimulation. Nucleus, apoptosis, and cytoskeleton staining of chondrocytes (a) and flow cytometry analysis (b) after IL-1β stimulation, scale bar = 50 μm. Expressions of apoptosis-related genes and proteins including Bax and Bcl2 (c) and ECM degeneration related genes and proteins including Adamts5 and MMP13 (d) from Control, Control + IL-1β, NAC + IL-1β, CH + IL-1β, and CH@Mn2+IL-1β groups. Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Control group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the Control + IL-1β group. ▽ p < 0.05, ▽▽ p < 0.01, and ▽▽▽ p < 0.001 suggests a significant difference in comparison to the NAC + IL-1β group.

Article Snippet: TUNEL staining was performed using a TUNEL Apoptosis Assay Kit (red fluorescence; Beyotime Biotechnology Company, China).

Techniques: Staining, Flow Cytometry, Control, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Schematic diagram illustrates the mechanism of Mn-NC SAzymes composite hydrogel inhibiting excessive activation of the MAPK signaling pathway, thereby suppressing inflammatory responses, extracellular matrix degradation, and cell apoptosis during the pathogenesis of TMJ-OA.

Article Snippet: TUNEL staining was performed using a TUNEL Apoptosis Assay Kit (red fluorescence; Beyotime Biotechnology Company, China).

Techniques: Activation Assay