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elisa kit  (Elabscience Biotechnology)


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    Elabscience Biotechnology elisa kit
    Elisa Kit, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/camp+elisa+kit/cAMP+(Cyclic+adenosine+monophosphate)+ELISA+Kit/10__1038_slash_s41467___026___77923___0-489-8-13
    Average 96 stars, based on 187 article reviews
    elisa kit - by Bioz Stars, 2026-10
    96/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Optimization of siRNA therapeutics targeting MIAT for cardioprotection in myocardial ischemia/reperfusion injury
    Article Snippet: .. Supernatants from cardiac tissue homogenates were collected by centrifugation at 5,000 × g for 10 min. cAMP levels in the supernatants were measured using a cAMP ELISA kit (Elabscience) according to the manufacturer’s protocol. ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Optimization of siRNA therapeutics targeting MIAT for cardioprotection in myocardial ischemia/reperfusion injury
    Article Snippet: .. Supernatants from cardiac tissue homogenates were collected by centrifugation at 5,000 × g for 10 min. cAMP levels in the supernatants were measured using a cAMP ELISA kit (Elabscience) according to the manufacturer’s protocol. ..

    Article Title: Discovery of kaempferol derivatives as novel PDE4 inhibitors for treatment of idiopathic pulmonary fibrosis.
    Article Snippet: Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal lung disease with a rapidly rising global incidence.. Despite its growing prevalence, effective therapeutic options for IPF remain limited.. Phosphodiesterase 4 (PDE4) inhibitors have recently emerged as promising anti-fibrotic candidates.

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment.
    Article Snippet: After twice washing with the blocking buffer and phosphate-buffered saline each, HRP signal was quantified by chemiluminescence with SuperSignal substrate (Vazyme, China) and an EnVisionTM Multilabel Plate Reader (PerkinElmer, USA). .. Quantification of intracellular cAMP levels by ELISA 24 h after transfection in the 6-well plate, LMH cell were re-passaged to a new 6-well plate at 1:2 ratio (for intracellular cAMP detection) and to a new 96-well plate at 1:30 ratio (for flag detection). after another 24 h, cells from each well of 6-well plate were digested for intracellular cAMP detection, which followed by instruction of cAMP ELISA Kit (E-EL-0056c, Elabscience, maximum concentration of standard:100 ng/L). .. After twice washing with the blocking buffer and phosphate-buffered saline each, HRP signal was quantified by chemiluminescence with SuperSignal substrate (Vazyme, China) and an EnVision Multilabel Plate Reader (PerkinElmer, USA).

    Article Title: Unraveling Cannabidiol's Dual Modulatory Role in Schizophrenia: Network Pharmacology and In Vivo Validation of Neuroinflammatory and Behavioral Modulation.
    Article Snippet: Schizophrenia (SCZ), a chronic psychiatric disorder, is characterized by cognitive impairment, hallucinations, and delusions, with current antipsychotic treatments offering limited efficacy and considerable side effects.. Cannabidiol (CBD), a nonpsychoactive compound from Cannabis sativa, has shown promise in treating neurological and psychiatric conditions, though its precise mechanisms in schizophrenia remain unclear.. Using network pharmacology, this study predicts CBD’s targets and pathways in schizophrenia, highlighting LPS-induced neuroinflammation and implicating 5-HT1AR-MAPK signaling as one potential contributor.

    Article Title: Unraveling Cannabidiol's Dual Modulatory Role in Schizophrenia: Network Pharmacology and In Vivo Validation of Neuroinflammatory and Behavioral Modulation
    Article Snippet: .. Intracellular cAMP levels were determined using a cAMP ELISA Kit (Elabscience®, cat. no. E-EL-0056) according to the manufacturer’s instructions. ..

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment
    Article Snippet: The tissue was homogenized on ice or in a refrigerated environment using RIPA lysis buffer (C0045, TargetMol, USA) supplemented with protease inhibitors (C0001, TargetMol, USA). .. Total protein concentration was determined using a BCA assay kit (Sigma-Aldrich, USA), followed by quantification of cAMP levels using a cAMP ELISA kit (E-EL-0056, Elabscience, China) according to the manufacturer’s protocol. .. 3T3-L1 cells were cultivated onto 6-well plates topped with a microscope slide (Thermo Scientific, 177399PK) and maintained in NCS media for 6 days.

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment.
    Article Snippet: after another 24 h, cells from each well of 6-well plate were digested for intracellular cAMP detection, which followed by instruction of cAMP ELISA Kit (E-EL-0056c, Elabscience, maximum concentration of standard:100 ng/L). .. Total protein concentration was determined using a BCA assay kit (Sigma‐Aldrich, USA), followed by quantification of cAMP levels using a cAMP ELISA kit (E-EL-0056, Elabscience, China) according to the manufacturer’s protocol. .. 3T3-L1 cell culture and differentiation 3T3-L1 cells were cultivated onto 6-well plates topped with a microscope slide (Thermo Scientific, 177399PK) and maintained in NCS media for 6 days.

    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
    Article Snippet: .. The intracellular cAMP level was examined by using a cAMP ELISA Kit (E-EL-0056, Elabscience, Wuhan, China) according to the manufacturer's instructions. .. The visualization of the docking results was done by using PyMOL (version 3.1.5.1, Schrödinger, Inc., New York, NY, USA) and Discovery Studio Visualizer (version 25.1.0.24284, BIOVIA, San Diego, CA, USA).

    Concentration Assay:

    Article Title: Discovery of kaempferol derivatives as novel PDE4 inhibitors for treatment of idiopathic pulmonary fibrosis.
    Article Snippet: Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal lung disease with a rapidly rising global incidence.. Despite its growing prevalence, effective therapeutic options for IPF remain limited.. Phosphodiesterase 4 (PDE4) inhibitors have recently emerged as promising anti-fibrotic candidates.

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment.
    Article Snippet: After twice washing with the blocking buffer and phosphate-buffered saline each, HRP signal was quantified by chemiluminescence with SuperSignal substrate (Vazyme, China) and an EnVisionTM Multilabel Plate Reader (PerkinElmer, USA). .. Quantification of intracellular cAMP levels by ELISA 24 h after transfection in the 6-well plate, LMH cell were re-passaged to a new 6-well plate at 1:2 ratio (for intracellular cAMP detection) and to a new 96-well plate at 1:30 ratio (for flag detection). after another 24 h, cells from each well of 6-well plate were digested for intracellular cAMP detection, which followed by instruction of cAMP ELISA Kit (E-EL-0056c, Elabscience, maximum concentration of standard:100 ng/L). .. After twice washing with the blocking buffer and phosphate-buffered saline each, HRP signal was quantified by chemiluminescence with SuperSignal substrate (Vazyme, China) and an EnVision Multilabel Plate Reader (PerkinElmer, USA).

    Transfection:

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment.
    Article Snippet: After twice washing with the blocking buffer and phosphate-buffered saline each, HRP signal was quantified by chemiluminescence with SuperSignal substrate (Vazyme, China) and an EnVisionTM Multilabel Plate Reader (PerkinElmer, USA). .. Quantification of intracellular cAMP levels by ELISA 24 h after transfection in the 6-well plate, LMH cell were re-passaged to a new 6-well plate at 1:2 ratio (for intracellular cAMP detection) and to a new 96-well plate at 1:30 ratio (for flag detection). after another 24 h, cells from each well of 6-well plate were digested for intracellular cAMP detection, which followed by instruction of cAMP ELISA Kit (E-EL-0056c, Elabscience, maximum concentration of standard:100 ng/L). .. After twice washing with the blocking buffer and phosphate-buffered saline each, HRP signal was quantified by chemiluminescence with SuperSignal substrate (Vazyme, China) and an EnVision Multilabel Plate Reader (PerkinElmer, USA).

    Expressing:

    Article Title: Unraveling Cannabidiol's Dual Modulatory Role in Schizophrenia: Network Pharmacology and In Vivo Validation of Neuroinflammatory and Behavioral Modulation.
    Article Snippet: Schizophrenia (SCZ), a chronic psychiatric disorder, is characterized by cognitive impairment, hallucinations, and delusions, with current antipsychotic treatments offering limited efficacy and considerable side effects.. Cannabidiol (CBD), a nonpsychoactive compound from Cannabis sativa, has shown promise in treating neurological and psychiatric conditions, though its precise mechanisms in schizophrenia remain unclear.. Using network pharmacology, this study predicts CBD’s targets and pathways in schizophrenia, highlighting LPS-induced neuroinflammation and implicating 5-HT1AR-MAPK signaling as one potential contributor.

    cAMP Assay:

    Article Title: Unraveling Cannabidiol's Dual Modulatory Role in Schizophrenia: Network Pharmacology and In Vivo Validation of Neuroinflammatory and Behavioral Modulation.
    Article Snippet: Schizophrenia (SCZ), a chronic psychiatric disorder, is characterized by cognitive impairment, hallucinations, and delusions, with current antipsychotic treatments offering limited efficacy and considerable side effects.. Cannabidiol (CBD), a nonpsychoactive compound from Cannabis sativa, has shown promise in treating neurological and psychiatric conditions, though its precise mechanisms in schizophrenia remain unclear.. Using network pharmacology, this study predicts CBD’s targets and pathways in schizophrenia, highlighting LPS-induced neuroinflammation and implicating 5-HT1AR-MAPK signaling as one potential contributor.

    Protein Concentration:

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment
    Article Snippet: The tissue was homogenized on ice or in a refrigerated environment using RIPA lysis buffer (C0045, TargetMol, USA) supplemented with protease inhibitors (C0001, TargetMol, USA). .. Total protein concentration was determined using a BCA assay kit (Sigma-Aldrich, USA), followed by quantification of cAMP levels using a cAMP ELISA kit (E-EL-0056, Elabscience, China) according to the manufacturer’s protocol. .. 3T3-L1 cells were cultivated onto 6-well plates topped with a microscope slide (Thermo Scientific, 177399PK) and maintained in NCS media for 6 days.

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment.
    Article Snippet: after another 24 h, cells from each well of 6-well plate were digested for intracellular cAMP detection, which followed by instruction of cAMP ELISA Kit (E-EL-0056c, Elabscience, maximum concentration of standard:100 ng/L). .. Total protein concentration was determined using a BCA assay kit (Sigma‐Aldrich, USA), followed by quantification of cAMP levels using a cAMP ELISA kit (E-EL-0056, Elabscience, China) according to the manufacturer’s protocol. .. 3T3-L1 cell culture and differentiation 3T3-L1 cells were cultivated onto 6-well plates topped with a microscope slide (Thermo Scientific, 177399PK) and maintained in NCS media for 6 days.

    BIA-KA:

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment
    Article Snippet: The tissue was homogenized on ice or in a refrigerated environment using RIPA lysis buffer (C0045, TargetMol, USA) supplemented with protease inhibitors (C0001, TargetMol, USA). .. Total protein concentration was determined using a BCA assay kit (Sigma-Aldrich, USA), followed by quantification of cAMP levels using a cAMP ELISA kit (E-EL-0056, Elabscience, China) according to the manufacturer’s protocol. .. 3T3-L1 cells were cultivated onto 6-well plates topped with a microscope slide (Thermo Scientific, 177399PK) and maintained in NCS media for 6 days.

    Article Title: Avian GCGR-mediated continuous fat utilization offers perspectives for obesity treatment.
    Article Snippet: after another 24 h, cells from each well of 6-well plate were digested for intracellular cAMP detection, which followed by instruction of cAMP ELISA Kit (E-EL-0056c, Elabscience, maximum concentration of standard:100 ng/L). .. Total protein concentration was determined using a BCA assay kit (Sigma‐Aldrich, USA), followed by quantification of cAMP levels using a cAMP ELISA kit (E-EL-0056, Elabscience, China) according to the manufacturer’s protocol. .. 3T3-L1 cell culture and differentiation 3T3-L1 cells were cultivated onto 6-well plates topped with a microscope slide (Thermo Scientific, 177399PK) and maintained in NCS media for 6 days.



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    Screening of the quinonoid compounds for the treatment of GIOP. (A) Flowchart depicting the screening process of the quinonoid compounds library. The schematic diagram was created by using BioRender.com. (B) Volcano diagram showing the effects of the 153 quinonoid compounds on Runx2 expression in BMSCs. Red and blue dots indicate the specific compounds that up- and down-regulate Runx2 expression in BMSCs, respectively. (C) Heat map showing the effect of the compounds on ALP activity in primary BMSCs. Color from blue to red indicates the ALP activity in primary BMSCs from low to high. (D) Measurement of <t>intracellular</t> ROS level in primary BMSCs treated with three potential compounds by using the fluorescent dye DCFDA. (E) Chemical structure of DUB, the final candidate among the screened drugs. (F) MTT assay for the proliferation of BMSCs treated with different doses of DUB for 2 and 10 days, under osteogenic induction conditions with or without 10 μM Dex. (G) Representative images and quantitative analysis of mineralized nodule formation via Alizarin Red S (ARS) staining in primary BMSCs treated with DUB at a series of concentrations, under osteogenic induction conditions with or without 10 μM Dex. (H) Western blot and quantification for the expression of osteogenesis-related proteins in primary BMSCs under different treatments. (I) Oil Red O staining and quantifications for lipid droplets 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 < 0.001 by one-way ANOVA. Scale bars: 200 μm (G), and 50 μm (I).
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    Image Search Results


    DMM reduced the IR-induced elevation of proinflammatory cytokine levels in BALF and SDH activity in lung tissue. (A) CINC-1 (B) IL-6, and (C) TNF-α levels in the BALF were measured using ELISA kits. In the IR group, levels of these cytokines were significantly higher compared to the control group. Treatment with 8 mM DMM markedly suppressed these increases following IR (D) SDH enzymatic activity was measured in lung tissue homogenates to evaluate target engagement. DMM significantly inhibited the IR-induced elevation of SDH activity. However, the anti-inflammatory effects and the inhibition of SDH activity of DMM were reversed when co-administered with 8 mM DMS. The measurement data are shown as the means ± SDs (n = 6 for each group). ***p < 0.001 indicate statistical significance compared with the control group, +++ p < 0.001 indicates statistical significance compared with the IR group, and ###p < 0.001 indicates statistical significance compared with the IR + DMM group.

    Journal: Frontiers in Pharmacology

    Article Title: Dimethyl malonate suppresses succinate dehydrogenase to alleviate ischemia/reperfusion-induced acute lung injury

    doi: 10.3389/fphar.2026.1746514

    Figure Lengend Snippet: DMM reduced the IR-induced elevation of proinflammatory cytokine levels in BALF and SDH activity in lung tissue. (A) CINC-1 (B) IL-6, and (C) TNF-α levels in the BALF were measured using ELISA kits. In the IR group, levels of these cytokines were significantly higher compared to the control group. Treatment with 8 mM DMM markedly suppressed these increases following IR (D) SDH enzymatic activity was measured in lung tissue homogenates to evaluate target engagement. DMM significantly inhibited the IR-induced elevation of SDH activity. However, the anti-inflammatory effects and the inhibition of SDH activity of DMM were reversed when co-administered with 8 mM DMS. The measurement data are shown as the means ± SDs (n = 6 for each group). ***p < 0.001 indicate statistical significance compared with the control group, +++ p < 0.001 indicates statistical significance compared with the IR group, and ###p < 0.001 indicates statistical significance compared with the IR + DMM group.

    Article Snippet: We utilized commercially available mouse ELISA kits (R&D Systems Inc., Minneapolis, MN, USA) to assess the concentrations of CINC-1, IL-6 and TNF-α in the BALF.

    Techniques: Activity Assay, Tissue, Enzyme-linked Immunosorbent Assay, Control, Drug discovery, Inhibition

    DMM attenuated HR-induced SDH activity, proinflammatory cytokine production and NF-κB signaling in MLE-12 cells. (A) The levels of KC/CXCL-1 in the cell culture supernatant were measured using ELISA (B) SDH enzymatic activity was measured in MLE-12 cell lysates to evaluate target engagement. DMM significantly inhibited the HR-induced elevation of SDH activity, which was reversed by DMS supplementation (C,D) Western blot analysis was used to evaluate p-IκB-α and IκB-α expression and p-NF-κB levels in MLE-12 cells subjected to 2 h of hypoxia followed by 1 h of reoxygenation. The measurement data are shown as the means ± SDs (n = 3 for each group). **p < 0.01 and ***p < 0.001 indicate statistical significance compared with the control group; + p < 0.05, and +++ p < 0.001 indicate statistical significance compared with the HR group; and ##p < 0.01 and ###p < 0.001 indicate statistical significance compared with the HR + DMM group.

    Journal: Frontiers in Pharmacology

    Article Title: Dimethyl malonate suppresses succinate dehydrogenase to alleviate ischemia/reperfusion-induced acute lung injury

    doi: 10.3389/fphar.2026.1746514

    Figure Lengend Snippet: DMM attenuated HR-induced SDH activity, proinflammatory cytokine production and NF-κB signaling in MLE-12 cells. (A) The levels of KC/CXCL-1 in the cell culture supernatant were measured using ELISA (B) SDH enzymatic activity was measured in MLE-12 cell lysates to evaluate target engagement. DMM significantly inhibited the HR-induced elevation of SDH activity, which was reversed by DMS supplementation (C,D) Western blot analysis was used to evaluate p-IκB-α and IκB-α expression and p-NF-κB levels in MLE-12 cells subjected to 2 h of hypoxia followed by 1 h of reoxygenation. The measurement data are shown as the means ± SDs (n = 3 for each group). **p < 0.01 and ***p < 0.001 indicate statistical significance compared with the control group; + p < 0.05, and +++ p < 0.001 indicate statistical significance compared with the HR group; and ##p < 0.01 and ###p < 0.001 indicate statistical significance compared with the HR + DMM group.

    Article Snippet: We utilized commercially available mouse ELISA kits (R&D Systems Inc., Minneapolis, MN, USA) to assess the concentrations of CINC-1, IL-6 and TNF-α in the BALF.

    Techniques: Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Drug discovery, Western Blot, Analysis, Expressing, Control

    Roles of A 2b R in ADO-mediated activation of the cAMP/PKA/CREB pathway in primary BMSCs. ( A ) Principal component analysis (PCA) of RNA-seq data from primary BMSCs treated with Dex or Dex + ADO. ( B ) The volcano plot presented the differentially expressed genes (DEGs) as determined by RNA-Seq in primary BMSCs treated with Dex or Dex + ADO. ( C ) Gene Ontology (GO) enrichment analysis in the biological process category for DEGs as determined by RNA-Seq in primary BMSCs treated with Dex, or Dex + ADO. ( D ) The molecular docking of ADO with mus musculus A 1 R, A 2a R, A 2b R, and A 3 R proteins. ADO is displayed in Cyan. The surrounding residues in the binding pocket are shown in green (forming a non-hydrogen bond with ADO) or magenta (forming a hydrogen bond with ADO). The hydrogen bond is labeled as yellow dashed lines. The backbone of the receptor is depicted as gray. ( E ) RT-qPCR analysis of the mRNA levels of Adora1 , Adora2a , Adora2b , and Adora3 in primary BMSCs treated with vehicle, Dex, or Dex + ADO. ( F ) RT-qPCR analysis for the expression of Runx2 in primary BMSCs of different groups. (G) Gene Set Enrichment Analysis (GSEA) plot showing the differentially expressed pathway (cAMP) between the Dex group and the Dex + ADO group as indicated by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. ( H ) Western blot validation for the knockdown deficiency of A 2b R after transfection with si Adora2b . ( I ) ELISA analysis for the relative intracellular cAMP levels in BMSCs of different groups. ( J ) Western blot and quantification for the expression of PKA, p-PKA, CREB, and p-CREB in primary BMSCs. ( K ) Representative images and quantitative analysis of Alizarin Red S staining for mineralization deposit in primary BMSCs of different groups under osteogenic conditions. n = 4 independent repeats by using different biological samples in each group for in vitro experiments. Data were means ± s.e.m. ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by one-way ANOVA. Scale bar: 200 μm (K).

    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 A 2b R in ADO-mediated activation of the cAMP/PKA/CREB pathway in primary BMSCs. ( A ) Principal component analysis (PCA) of RNA-seq data from primary BMSCs treated with Dex or Dex + ADO. ( B ) The volcano plot presented the differentially expressed genes (DEGs) as determined by RNA-Seq in primary BMSCs treated with Dex or Dex + ADO. ( C ) Gene Ontology (GO) enrichment analysis in the biological process category for DEGs as determined by RNA-Seq in primary BMSCs treated with Dex, or Dex + ADO. ( D ) The molecular docking of ADO with mus musculus A 1 R, A 2a R, A 2b R, and A 3 R proteins. ADO is displayed in Cyan. The surrounding residues in the binding pocket are shown in green (forming a non-hydrogen bond with ADO) or magenta (forming a hydrogen bond with ADO). The hydrogen bond is labeled as yellow dashed lines. The backbone of the receptor is depicted as gray. ( E ) RT-qPCR analysis of the mRNA levels of Adora1 , Adora2a , Adora2b , and Adora3 in primary BMSCs treated with vehicle, Dex, or Dex + ADO. ( F ) RT-qPCR analysis for the expression of Runx2 in primary BMSCs of different groups. (G) Gene Set Enrichment Analysis (GSEA) plot showing the differentially expressed pathway (cAMP) between the Dex group and the Dex + ADO group as indicated by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. ( H ) Western blot validation for the knockdown deficiency of A 2b R after transfection with si Adora2b . ( I ) ELISA analysis for the relative intracellular cAMP levels in BMSCs of different groups. ( J ) Western blot and quantification for the expression of PKA, p-PKA, CREB, and p-CREB in primary BMSCs. ( K ) Representative images and quantitative analysis of Alizarin Red S staining for mineralization deposit in primary BMSCs of different groups under osteogenic conditions. n = 4 independent repeats by using different biological samples in each group for in vitro experiments. Data were means ± s.e.m. ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by one-way ANOVA. Scale bar: 200 μm (K).

    Article Snippet: The intracellular cAMP level was examined by using a cAMP ELISA Kit (E-EL-0056, Elabscience, Wuhan, China) according to the manufacturer's instructions.

    Techniques: Activation Assay, RNA Sequencing, Binding Assay, Labeling, Quantitative RT-PCR, Expressing, Western Blot, Biomarker Discovery, Knockdown, Transfection, Enzyme-linked Immunosorbent Assay, Staining, In Vitro

    Screening of the quinonoid compounds for the treatment of GIOP. (A) Flowchart depicting the screening process of the quinonoid compounds library. The schematic diagram was created by using BioRender.com. (B) Volcano diagram showing the effects of the 153 quinonoid compounds on Runx2 expression in BMSCs. Red and blue dots indicate the specific compounds that up- and down-regulate Runx2 expression in BMSCs, respectively. (C) Heat map showing the effect of the compounds on ALP activity in primary BMSCs. Color from blue to red indicates the ALP activity in primary BMSCs from low to high. (D) Measurement of intracellular ROS level in primary BMSCs treated with three potential compounds by using the fluorescent dye DCFDA. (E) Chemical structure of DUB, the final candidate among the screened drugs. (F) MTT assay for the proliferation of BMSCs treated with different doses of DUB for 2 and 10 days, under osteogenic induction conditions with or without 10 μM Dex. (G) Representative images and quantitative analysis of mineralized nodule formation via Alizarin Red S (ARS) staining in primary BMSCs treated with DUB at a series of concentrations, under osteogenic induction conditions with or without 10 μM Dex. (H) Western blot and quantification for the expression of osteogenesis-related proteins in primary BMSCs under different treatments. (I) Oil Red O staining and quantifications for lipid droplets 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 < 0.001 by one-way ANOVA. Scale bars: 200 μm (G), and 50 μm (I).

    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: Screening of the quinonoid compounds for the treatment of GIOP. (A) Flowchart depicting the screening process of the quinonoid compounds library. The schematic diagram was created by using BioRender.com. (B) Volcano diagram showing the effects of the 153 quinonoid compounds on Runx2 expression in BMSCs. Red and blue dots indicate the specific compounds that up- and down-regulate Runx2 expression in BMSCs, respectively. (C) Heat map showing the effect of the compounds on ALP activity in primary BMSCs. Color from blue to red indicates the ALP activity in primary BMSCs from low to high. (D) Measurement of intracellular ROS level in primary BMSCs treated with three potential compounds by using the fluorescent dye DCFDA. (E) Chemical structure of DUB, the final candidate among the screened drugs. (F) MTT assay for the proliferation of BMSCs treated with different doses of DUB for 2 and 10 days, under osteogenic induction conditions with or without 10 μM Dex. (G) Representative images and quantitative analysis of mineralized nodule formation via Alizarin Red S (ARS) staining in primary BMSCs treated with DUB at a series of concentrations, under osteogenic induction conditions with or without 10 μM Dex. (H) Western blot and quantification for the expression of osteogenesis-related proteins in primary BMSCs under different treatments. (I) Oil Red O staining and quantifications for lipid droplets 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 < 0.001 by one-way ANOVA. Scale bars: 200 μm (G), and 50 μm (I).

    Article Snippet: The intracellular cAMP level was examined by using a cAMP ELISA Kit (E-EL-0056, Elabscience, Wuhan, China) according to the manufacturer's instructions.

    Techniques: Expressing, Activity Assay, MTT Assay, Staining, Western Blot, In Vitro

    Roles of A 2b R in ADO-mediated activation of the cAMP/PKA/CREB pathway in primary BMSCs. ( A ) Principal component analysis (PCA) of RNA-seq data from primary BMSCs treated with Dex or Dex + ADO. ( B ) The volcano plot presented the differentially expressed genes (DEGs) as determined by RNA-Seq in primary BMSCs treated with Dex or Dex + ADO. ( C ) Gene Ontology (GO) enrichment analysis in the biological process category for DEGs as determined by RNA-Seq in primary BMSCs treated with Dex, or Dex + ADO. ( D ) The molecular docking of ADO with mus musculus A 1 R, A 2a R, A 2b R, and A 3 R proteins. ADO is displayed in Cyan. The surrounding residues in the binding pocket are shown in green (forming a non-hydrogen bond with ADO) or magenta (forming a hydrogen bond with ADO). The hydrogen bond is labeled as yellow dashed lines. The backbone of the receptor is depicted as gray. ( E ) RT-qPCR analysis of the mRNA levels of Adora1 , Adora2a , Adora2b , and Adora3 in primary BMSCs treated with vehicle, Dex, or Dex + ADO. ( F ) RT-qPCR analysis for the expression of Runx2 in primary BMSCs of different groups. (G) Gene Set Enrichment Analysis (GSEA) plot showing the differentially expressed pathway (cAMP) between the Dex group and the Dex + ADO group as indicated by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. ( H ) Western blot validation for the knockdown deficiency of A 2b R after transfection with si Adora2b . ( I ) ELISA analysis for the relative intracellular cAMP levels in BMSCs of different groups. ( J ) Western blot and quantification for the expression of PKA, p-PKA, CREB, and p-CREB in primary BMSCs. ( K ) Representative images and quantitative analysis of Alizarin Red S staining for mineralization deposit in primary BMSCs of different groups under osteogenic conditions. n = 4 independent repeats by using different biological samples in each group for in vitro experiments. Data were means ± s.e.m. ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by one-way ANOVA. Scale bar: 200 μm (K).

    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 A 2b R in ADO-mediated activation of the cAMP/PKA/CREB pathway in primary BMSCs. ( A ) Principal component analysis (PCA) of RNA-seq data from primary BMSCs treated with Dex or Dex + ADO. ( B ) The volcano plot presented the differentially expressed genes (DEGs) as determined by RNA-Seq in primary BMSCs treated with Dex or Dex + ADO. ( C ) Gene Ontology (GO) enrichment analysis in the biological process category for DEGs as determined by RNA-Seq in primary BMSCs treated with Dex, or Dex + ADO. ( D ) The molecular docking of ADO with mus musculus A 1 R, A 2a R, A 2b R, and A 3 R proteins. ADO is displayed in Cyan. The surrounding residues in the binding pocket are shown in green (forming a non-hydrogen bond with ADO) or magenta (forming a hydrogen bond with ADO). The hydrogen bond is labeled as yellow dashed lines. The backbone of the receptor is depicted as gray. ( E ) RT-qPCR analysis of the mRNA levels of Adora1 , Adora2a , Adora2b , and Adora3 in primary BMSCs treated with vehicle, Dex, or Dex + ADO. ( F ) RT-qPCR analysis for the expression of Runx2 in primary BMSCs of different groups. (G) Gene Set Enrichment Analysis (GSEA) plot showing the differentially expressed pathway (cAMP) between the Dex group and the Dex + ADO group as indicated by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. ( H ) Western blot validation for the knockdown deficiency of A 2b R after transfection with si Adora2b . ( I ) ELISA analysis for the relative intracellular cAMP levels in BMSCs of different groups. ( J ) Western blot and quantification for the expression of PKA, p-PKA, CREB, and p-CREB in primary BMSCs. ( K ) Representative images and quantitative analysis of Alizarin Red S staining for mineralization deposit in primary BMSCs of different groups under osteogenic conditions. n = 4 independent repeats by using different biological samples in each group for in vitro experiments. Data were means ± s.e.m. ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by one-way ANOVA. Scale bar: 200 μm (K).

    Article Snippet: The intracellular cAMP level was examined by using a cAMP ELISA Kit (E-EL-0056, Elabscience, Wuhan, China) according to the manufacturer's instructions.

    Techniques: Activation Assay, RNA Sequencing, Binding Assay, Labeling, Quantitative RT-PCR, Expressing, Western Blot, Biomarker Discovery, Knockdown, Transfection, Enzyme-linked Immunosorbent Assay, Staining, In Vitro