one step tunel apoptosis 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: 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: 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: Frontiers in Veterinary Science

Article Title: Chitosan-selenium alleviates AFB1-induced growth impairment and immunotoxicity in broilers by suppressing splenic oxidative stress, inflammation and apoptosis

doi: 10.3389/fvets.2026.1859938

Figure Lengend Snippet: Ct-Se alleviated AFB1-induced histopathological damage in spleen. Histological examination of H&E staining of the spleen of broilers in the CON (A) , AFB1 (B) , Ct-Se (C) , and AFB1 + Ct-Se (D) groups, respectively, ( n = 3). The scale bar represents 100 μm, △ indicates splenic nodules. In the AFB1 group, the boundaries between red and white pulp were indistinct, red pulp congestion (blue arrows), lymphocyte reduction (white arrows) and apoptosis (yellow arrows) were observed. The CON and Ct-Se groups maintained normal tissue structure, and co-treatment with Ct-Se notably improved histological architecture.

Article Snippet: Briefly, sections were treated with proteinase K, and subsequently stained using a one-step TUNEL apoptosis assay kit (Jiangsu KeyGen Biotech Co., Ltd., China) according to the manufacturer’s instructions.

Techniques: Staining

Journal: Frontiers in Veterinary Science

Article Title: Chitosan-selenium alleviates AFB1-induced growth impairment and immunotoxicity in broilers by suppressing splenic oxidative stress, inflammation and apoptosis

doi: 10.3389/fvets.2026.1859938

Figure Lengend Snippet: Ct-Se attenuated AFB1-induced apoptosis in spleen determined by TUNEL staining. (A) Representative the TUNEL staining results in spleen sections ( n = 3). The TUNEL-positive cells were stained green, and the bars represent 50 μm. (B) Quantification of apoptosis index using Image J. The formula used for assessing the apoptosis rate was the number of TUNEL-positive nuclei (TUNEL specks) × 100%/the total number of nuclei (DAPI), and the data are expressed as mean ± SD (n = 6). ** p < 0.01 indicates significant difference compared to the CON group; ## p < 0.01 < # p < 0.05 indicates significant difference compared to the AFB1 group.

Article Snippet: Briefly, sections were treated with proteinase K, and subsequently stained using a one-step TUNEL apoptosis assay kit (Jiangsu KeyGen Biotech Co., Ltd., China) according to the manufacturer’s instructions.

Techniques: TUNEL Assay, Staining

Journal: Frontiers in Veterinary Science

Article Title: Chitosan-selenium alleviates AFB1-induced growth impairment and immunotoxicity in broilers by suppressing splenic oxidative stress, inflammation and apoptosis

doi: 10.3389/fvets.2026.1859938

Figure Lengend Snippet: Ct-Se attenuated AFB1-induced apoptosis-related protein expression in spleen of broilers. (A) Western blot analysis of apoptosis-related proteins in spleen tissue. (B–F) Relative protein expression levels of Bcl-2, Bax, Bax/Bcl-2, cleaved-caspase-9, and cleaved-caspase-3. Data are presented as mean ± SD ( n = 6). ** p < 0.01 < * p < 0.05 indicates significant difference compared to the CON group; ## p < 0.01 < # p < 0.05 indicates significant difference compared to the AFB1 group; ‡ p < 0.01 < † p < 0.05 indicates significant difference between the Ct-Se group and the AFB1 + Ct-Se group.

Article Snippet: Briefly, sections were treated with proteinase K, and subsequently stained using a one-step TUNEL apoptosis assay kit (Jiangsu KeyGen Biotech Co., Ltd., China) according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot