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: 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

Journal: Poultry Science

Article Title: NRF2 deficiency impairs proliferation and survival of chicken primordial germ cells via oxidative stress, mitochondrial dysfunction and apoptosis

doi: 10.1016/j.psj.2026.106765

Figure Lengend Snippet: Inhibition of NRF2 results in reduced proliferation and increased apoptosis in chicken PGCs. (A) Morphological observation of PGCs treated with varying concentrations of ML385. Scale bar: 50 μm. (B) Comparison of cell numbers after 3 d of culture in media containing different concentrations of ML385. Different lowercase letters indicate significant differences among groups ( p < 0.05). Based on these dose-dependent effects, the 24 μM concentration was selected for all subsequent experiments. (C and D) Analysis of cell proliferation capacity via EdU assay in ML385-PGCs and Ctrl-PGCs. Scale bar: 50 μm. (E-G) Evaluation of apoptosis levels using Annexin V and PI staining in ML385-PGCs and Ctrl-PGCs. (H and I) Detection of NRF2 protein expression in ML385-PGCs and Ctrl-PGCs by Western blotting. All data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance is indicated by * ( p < 0.05) and *** ( p < 0.001).

Article Snippet: Apoptosis was assessed using the Annexin V‐FITC/PI Apoptosis Detection Kit (40302ES50, Yeasen, Shanghai, China) according to the manufacturer's instructions.

Techniques: Inhibition, Comparison, Concentration Assay, EdU Assay, Staining, Expressing, Western Blot

Journal: Poultry Science

Article Title: NRF2 deficiency impairs proliferation and survival of chicken primordial germ cells via oxidative stress, mitochondrial dysfunction and apoptosis

doi: 10.1016/j.psj.2026.106765

Figure Lengend Snippet: RNA sequencing analysis reveals aberrant expression of genes related to cell growth, cell cycle, and apoptosis induced by NRF2 inhibition. GSEA of genes related to cell growth (A), cell cycle (B), and apoptosis (C and D) in ML385-PGCs and Ctrl-PGCs. Heatmap (E) depicting the expression of apoptosis pathway genes between ML385-PGCs and Ctrl-PGCs. Comparative analysis (F) of normalized FPKM values of apoptosis pathway genes between ML385-PGCs and Ctrl-PGCs. Statistical significance is indicated by * ( p < 0.05).

Article Snippet: Apoptosis was assessed using the Annexin V‐FITC/PI Apoptosis Detection Kit (40302ES50, Yeasen, Shanghai, China) according to the manufacturer's instructions.

Techniques: RNA Sequencing, Expressing, Inhibition

Journal: Poultry Science

Article Title: NRF2 deficiency impairs proliferation and survival of chicken primordial germ cells via oxidative stress, mitochondrial dysfunction and apoptosis

doi: 10.1016/j.psj.2026.106765

Figure Lengend Snippet: Schematic representation illustrating the impact of NRF2 deficiency on chicken PGCs. NRF2 plays a critical role in maintaining the normal growth of PGCs during in vitro culture by activating antioxidant pathways. NRF2 deficiency impairs antioxidant defense, leading to oxidative stress, mitochondrial damage, and increased apoptosis. Additionally, mitophagy and autophagy are activated to remove damaged organelles. Furthermore, elevated ferroptosis markers, including oxidative stress‑induced lipid peroxidation and Fe²⁺ accumulation, are observed, contributing to reduced PGC viability. Δψm denotes mitochondrial membrane potential.

Article Snippet: Apoptosis was assessed using the Annexin V‐FITC/PI Apoptosis Detection Kit (40302ES50, Yeasen, Shanghai, China) according to the manufacturer's instructions.

Techniques: In Vitro, Membrane

Journal: International Journal of Pharmaceutics: X

Article Title: A pH-responsive dual-drug nanoplatform for stromal remodeling and enhanced chemotherapy via MMP3/TGF- β inhibition

doi: 10.1016/j.ijpx.2026.100489

Figure Lengend Snippet: In vitro therapeutic efficacy and synergy analysis of RPAE-QM in 4 T1 cells. (A) Cell viability of 4 T1 cells incubated with different formulations for 48 h determined by MTT assay. (B) The corresponding IC 50 values of the respective treatments. (C) Representative flow cytometry plots of Annexin V-FITC/PI staining for apoptosis analysis in 4 T1 cells. (D) Quantitative analysis of the total apoptotic rate. (E) Dose-response curves of free Que., free DM1, and their combination used for quantitative synergy determination. (F) The Combination Index (CI) plot as a function of Fraction affected (Fa) generated using the Chou-Talalay method; the reference line at CI = 1 indicates an additive effect, while CI < 1 indicates synergism. Data are presented as mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.

Article Snippet: After 24 h, the cells were harvested and stained with Annexin V-FITC/PI Cell Apoptosis Detection Kit (Beyotime, China) for flow cytometer analysis.

Techniques: In Vitro, Drug discovery, Incubation, MTT Assay, Flow Cytometry, Staining, Generated

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