elisa kits for pge2  (Elabscience Biotechnology)

Journal: Bioactive Materials

Article Title: Mesenchymal stromal cells-loaded 3D radially aligned composite scaffold with potentiated paracrine signaling for sequential bone regeneration

doi: 10.1016/j.bioactmat.2026.02.059

Figure Lengend Snippet: Temporal analysis of the BMSC paracrine profile on different scaffolds. (A) Confocal microscopy images from Live/Dead fluorescence staining of BMSCs encapsulated within the PCL/HAp-GelMA/BMSCs scaffold after 1, 3, 5, and 14 d of 3D culture (live cells, green; dead cells, red). (B) The concentrations of key paracrine factors (TGF-β, PGE2, VEGF, HGF, and BMP-2) from BMSCs cultured in different scaffolds, quantified from culture supernatants at day 3 and day 7. (C) Corresponding relative mRNA expression levels of TGFB1, PTGS2, VEGFA, HGF, and BMP-2 in BMSCs at day 3 and day 7, as determined by qPCR analysis. Data are presented as mean ± SD (n = 3) *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns: not significant.

Article Snippet: ELISA kits for PGE2 (Cat. No. E-EL-0034), TGF-β (Cat. No. E-EL-0162), VEGF (Cat. No. E-EL-R2603), and HGF (Cat. No. E-EL-R0496) were purchased from Elabscience (Wuhan, China).

Techniques: Confocal Microscopy, Fluorescence, Staining, Cell Culture, Expressing

Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: SCS reprograms the lineage commitment of MSCs after GC treatment and inhibits the generation of primary senescent adipocytes. ( A ) Schematic illustration of the in vitro investigation of SCS targeting the prostaglandin/PPARγ/INK positive feedback loop in MPS-induced primary senescent adipocytes. ( B ) Representative flow cytometry plot showing p16 + senescent cells in adipocytes derived from bone marrow after 14 days of in vivo MPS induction and subsequently treated with SCS in vitro . ( C ) qPCR analysis of 12 senescence-associated markers in primary senescent adipocytes after in vitro SCS treatment. n = 3 biological replicates. ( D ) ELISA analysis of IL-1β levels in adipocyte supernatant following in vitro SCS treatment. n = 6 biological replicates. ( E ) ELISA analysis of secreted prostaglandins PGD2 and PGE2 in adipocytes under different treatment conditions. D-PBS: bone marrow adipocytes isolated from mice treated in vivo with the solvent control DMSO, followed by in vitro treatment with PBS; M-PBS: bone marrow adipocytes isolated from mice treated in vivo with MPS, followed by in vitro treatment with PBS. M-SCS: bone marrow adipocytes isolated from mice treated in vivo with MPS, followed by in vitro treatment with SCS. ( F ) Western blot analysis of intracellular COX-2 protein levels in adipocytes across the three treatment conditions. ( G ) Schematic illustration of competitive osteogenic–adipogenic differentiation of CD45 − Ter119 − CD31 − LepR + MSCs after 7 days of in vivo SCS and MPS co-treatment. ( H ) qPCR analysis of pan-adipocyte markers ( Fabp4 , Adipoq , Plin1 , Cd36 , and Lep ) in CD45 − Ter119 − CD31 − LepR + MSCs after 14 days of in vitro competitive lineage differentiation. n = 3 biological replicates. ( I and J ) Representative immunofluorescence images (I) and quantification (J) of perilipin + adipocytes and osteopontin + mature osteoblasts derived from lineage-committed MSCs. n = 6 biological replicates. (Scale bars, 30 μm, 15 μm and 15 μm). ( K ) Western blot analysis of adipogenesis-related markers C/EBPα, PPARγ, and C/EBPβ in the lineage-mixed cells after in vitro competitive differentiation of CD45 − Ter119 − CD31 − LepR + MSCs. ( L ) qPCR analysis of lipogenesis-related markers Fasn , Scd1 , Srebf1 , Acaca , and Acacb . n = 3 biological replicates. ( M and N ) Representative H&E staining images (M) of the femurs at day 14 following SCS and MPS co-treatment. Yellow arrows indicate bone marrow adipocytes. Magnified images show hypertrophic adipocyte morphology, with quantification of adipocyte diameter (N). n = 19 biological replicates. (Scale bars, 200 μm, 50 μm and 20 μm). Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant. Statistical significance was determined using an unpaired two-tailed Student's t -test ( C, D, H, J, L and N ), or one-way ANOVA with Tukey's post hoc test ( E ).

Article Snippet: To further investigate the anti-senescent effects of SCS on primary senescent adipocytes, ELISA was performed on adipocyte supernatant to quantify senescence-associated factors including IL-1β (Neobioscience, EMC001b.96), IL-18 (Neobioscience, EMC011.96), TNF-α (Neobioscience, EMC102a.96), and prostaglandins PGJ2 (NOVUS, NBP2-61285), PGD2 (Cayman Chemical, 500151), and PGE2 (R&D Systems, KGE004B).

Techniques: In Vitro, Flow Cytometry, Derivative Assay, In Vivo, Enzyme-linked Immunosorbent Assay, Isolation, Solvent, Control, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Interleukin-1β-induced arthritis involves chondrocyte oxiapoptophagy

doi: 10.4196/kjpp.25.279

Figure Lengend Snippet: (A, B) IL-1β increases ROS production in chondrocytes. Scale bar = 100 μm. (C–E) The expression of inflammatory mediators, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nitric oxide (NO), and prostaglandin E 2 (PGE 2 ), are upregulated in chondrocytes stimulated with IL-1β. Values are presented as mean ± SD. DCF-DA, dichlorfluorescein-diacetate.

Article Snippet: The production of PGE 2 in the chondrocytes stimulated with IL-1β was measured using a PGE 2 Parameter Assay Kit (R&D System), according to the manufacturer’s instruction.

Techniques: Expressing

Journal: Cancer Research

Article Title: ACSL5 Mediates Adaptation to the Palmitic Acid–Enriched Pulmonary Microenvironment to Enhance Metastatic Breast Cancer Cell Survival and Lung Metastasis

doi: 10.1158/0008-5472.CAN-25-0866

Figure Lengend Snippet: PA-ACSL5 induced COX2 expression and PGE2 accumulation to promote survival and growth of lung metastatic breast cancer cells. A, Expression heatmap of genes upregulated in the control and ACSL5-knockdown LM3 cells treated with or without PA. B, Kaplan–Meier survival curves for lung metastasis–free survival of patients with breast cancer with low and high COX2 mRNA levels in the lung metastases ( GSE5327 ). C, PGE2 levels in the lung metastases from patients with breast cancer compared with the control lung tissue. D, The control and ACSL5-knockdown LM3 cells were treated with PA. In the case of PA adaptation, Western blotting was used to test the protein expression in the indicated groups. E, ELISA assay was used to evaluate the PGE2 levels in the indicated group. F–J, ACSL5-silenced LM3 cells were transfected with ectopic COX2 or treated with PGE2 (10 µmol/L). Flow cytometry (Annexin V–APC/PI staining; F and G ), colony formation ( H and I ), and proliferation ( J ) in the indicated groups are shown. All data represent the mean ± SD. Log-rank test ( B ), Mann–Whitney test ( C ), one-way ANOVA ( E , G , and I ), and two-way ANOVA ( J ) were utilized. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Lung interstitial fluid and culture medium were collected and analyzed using the Prostaglandin E2 Assay Kit (R&D Systems) according to the manufacturer’s instructions.

Techniques: Expressing, Control, Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay, Transfection, Flow Cytometry, Staining, MANN-WHITNEY

Journal: Cancer Research

Article Title: ACSL5 Mediates Adaptation to the Palmitic Acid–Enriched Pulmonary Microenvironment to Enhance Metastatic Breast Cancer Cell Survival and Lung Metastasis

doi: 10.1158/0008-5472.CAN-25-0866

Figure Lengend Snippet: PA-ACSL5 induced COX2 expression and PGE2 accumulation activates EP4 to foster DTC survival and proliferation through PI3K/AKT and ERK signaling. A, EP4 mRNA levels in clinical lung, liver, brain, and bone metastases of patients with breast cancer (Met500). B and C, LM3 cells were treated with PGE2 alone or a combination of PGE2 and EP1-4 inhibitors (EP1/2i, EP3i, and EP4i). Annexin V–APC/PI staining ( B ) and proliferation assay ( C ) were performed. D and E, Activation of PI3K/AKT, ERK, and β-catenin signaling was detected in indicated groups. F, Western blotting was used to evaluate the activation of PI3K/AKT and ERK signaling in the indicated LM3 cells with shNC, shEP4#1, shEP4#2, and shNC + ONO-AE3-208. G, ACSL5-knockdown LM3 cells were transfected with ectopic COX2 or treated with PGE2 (left), or ACSL5-overexpressed primary tumor cells (PRI) were transfected with shCOX2 (right). Activation of PI3K/AKT and ERK signaling was detected. H, The indicated engineered MDA-MB-231/LM3 cells (2 × 10 6 /mouse) were injected into BALB/c nude mice, and the metastatic burden of each group was assessed by BLI. All data represent the mean ± SD. One-way ANOVA ( A , B , and H ) and two-way ANOVA ( C ) were used. **, P < 0.01; ***, P < 0.001; ns, not significant.

Article Snippet: Lung interstitial fluid and culture medium were collected and analyzed using the Prostaglandin E2 Assay Kit (R&D Systems) according to the manufacturer’s instructions.

Techniques: Expressing, Staining, Proliferation Assay, Activation Assay, Western Blot, Knockdown, Transfection, Injection