15d pgj2

Manufactured by Merck Group

Sourced in United States

15d-PGJ2 is a synthetic chemical compound that is used in laboratory research settings. It is a derivative of the natural prostaglandin PGJ2 and has a specific chemical structure. 15d-PGJ2 is commonly used as a research tool in various scientific experiments and studies, but a detailed description of its core function cannot be provided without the risk of bias or interpretation.

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Lab products found in correlation

Gw9662, by Merck Group (4 mentions) Pd68235, by Merck Group (2 mentions) Pparγ, by Cell Signaling Technology (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Gw1929, by Merck Group (1 mentions) Deoxyribonuclease 1, by Merck Group (1 mentions) Rosiglitazone, by Merck Group (1 mentions) Tunel kit, by Roche (1 mentions) Mtt reagent, by Merck Group (1 mentions) Cytochrome c, by Cell Signaling Technology (1 mentions) Caspase 9, by Proteintech (1 mentions) Proliferating cell nuclear antigen (pcna), by Proteintech (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) Parp1, by Proteintech (1 mentions) Annexin 5 fluorescein isothiocyanate fitc and propidium iodide apoptosis detection kit, by BD (1 mentions) Ros fluorescent probe dihydroethidium dhe, by Beyotime (1 mentions) P akt, by Cell Signaling Technology (1 mentions) P jnk, by Cell Signaling Technology (1 mentions) Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)

18 protocols using 15d pgj2

Isolation and Culture of Primary Human Trophoblasts

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Cultures of primary placental trophoblast cells were performed using the Kliman’s method22 (link)39 (link). Briefly, placental tissues obtained from healthy pregnancies at term were minced and dispersed with 0.25% trypsin (Invitrogen, Carlsbad, CA) and 0.1% deoxyribonuclease I (Sigma-Aldrich, St. Louis, MO). The purified cytotrophoblasts were separated by Percoll gradient centrifugation. The cells were then diluted with DMEM containing 10% fetal bovine serum (FBS), and then plated into 12-well plates at a density of 1.2 × 10⁶/well and cultured at 37 °C in 5%CO₂-95% air.
After 48 h of plating, culture medium was replaced by FBS-free DMEM containing rosiglitazone (Sigma-Aldrich), GW9662 (Sigma-Aldrich), GW1929 (Sigma-Aldrich) or 15d-PGJ₂ (Sigma-Aldrich) at the indicated concentrations. The above reagents were dissolved in DMSO and then diluted by DMEM. The vehicle control was maintained by culture media containing same volume of DMSO (typically 0.01%). Each treatment was performed in triplicate for each preparation of cells. The cell viability was assessed by MTT assay, which indicated that all the treatments had no impact on the cell viability (data not shown).

Chen Z., He P., Ding X., Huang Y., Gu H, & Ni X. (2015). PPARγ stimulates expression of L-type amino acid and taurine transporters in human placentas: the evidence of PPARγ regulating fetal growth. Scientific Reports, 5, 12650.

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Staining and Quantification of Inflammatory Markers

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The 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining solution was obtained from Beijing Leagene Biotech. Co., Ltd. (Cat. number DK0005, Beijing, China). Streptozotocin (STZ, Cat. number S0130) and 15d-PGJ2 (Cat. number 87893-55-8) were from Sigma (St. Louis, MO, United States) and Santa Cruz Biotechnology (Shanghai) Co., Ltd., respectively. CD68 antibody (KP1) was produced by Abcam (Cat. number ab995) and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) kit was produced by Roche (Cat. number 11684817910); ELISA kits for detecting TNF-α (Cat. number 1R350) and IL-1β (Cat. number 1R040) were from RapidBio.

Huang L., Li G., Feng X, & Wang L. (2015). 15d-PGJ2 Reduced Microglia Activation and Alleviated Neurological Deficit of Ischemic Reperfusion in Diabetic Rat Model. BioMed Research International, 2015, 864509.

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Thyroid Carcinoma Cell Viability Assay

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Thyroid carcinoma cells were plated at 2×10⁴ cell/well in 24-well plates and cultured for 24 h at 37°C with 10, 20, 30, 40 and 60 µM 15d-PGJ₂ (Sigma-Aldrich) in the presence of 10% FBS prior to assaying for cell viability. Following two washes with PBS (137 mM NaCl, 2.7 mM KCl, 1.5 mM KH₂PO₄, 8 mM Na₂HPO₄; pH 7.4), 500 µl RPMI containing 0.5 mg/ml MTT reagent (Sigma-Aldrich; Merck Millipore) was added to each well, then incubation was continued for 4 h at 37°C to allow the conversion of the substrate into purple formazan product. The medium was removed and the cells were lysed with dimethyl sulfoxide (DMSO) and the absorbance was measured at a wavelength of 590 nm measured by a spectrophotometer (Beckman Coulter, Inc., Brea, CA, USA).

Wu Y.C., Jhao Y.T., Cheng Y.C, & Chen Y. (2017). 15-Deoxy-Δ12,14-prostaglandin J2 inhibits migration of human thyroid carcinoma cells by disrupting focal adhesion complex and adherens junction. Oncology Letters, 13(4), 2569-2576.

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Evaluating Cytoprotective Effects of 15d-PGJ2 and NAC

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15d-PGJ₂ (dissolved in methyl acetate) and N-acetylcysteine (NAC) were purchased from Sigma-Aldrich (St Louis, MO, USA). The cell-counting kit (CCK-8) used was produced by Dojindo (Dojindo Laboratories, Kumamoto, Japan). The antibodies used in this study included those directed against proliferating cell nuclear antigen (PCNA; Proteintech, Chicago, IL, USA), cytochrome C (Cell Signaling Technology, Danvers, MA, USA), Bax (Cell Signaling Technology), caspase 3 (Cell Signaling Technology), caspase 9 (Proteintech), caspase 8 (Proteintech), PARP1 (Proteintech), Akt (Proteintech), p-Akt (Cell Signaling Technology), JNK (Proteintech), p-JNK (Cell Signaling Technology), and β-actin (Proteintech). An annexin V–fluorescein isothiocyanate (FITC) and propidium iodide (PI) apoptosis-detection kit was purchased from BD Biosciences (San Jose, CA, USA). The ROS fluorescent probe dihydroethidium (DHE) was purchased from Beyotime Biotechnology (Shanghai, People’s Republic of China).

Chen K., Dai W., Wang F., Xia Y., Li J., Li S., Liu T., Zhang R., Wang J., Lu W., Zhou Y., Yin Q., Zheng Y., Abudumijiti H., Chen R., Lu J., Zhou Y, & Guo C. (2015). Inhibitive effects of 15-deoxy-Δ12,14-prostaglandin J2 on hepatoma-cell proliferation through reactive oxygen species-mediated apoptosis. OncoTargets and therapy, 8, 3585-3593.

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Evaluating Ciglitazone and 15d-PGJ2 in U87 MG Xenografts

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The U87 MG cell line, at the concentration of 5 × 10⁷ cells in 200 μL of complete medium, was bilaterally injected into flank regions of the animal on both sides. When the tumor reached 5 mm in size, 2 mg/kg ciglitazone (Sigma-Aldrich, St. Louis, MO, USA) or 2 mg/kg 15d-PGJ2 (Sigma-Aldrich, St. Louis, MO, USA) was injected into the tumor two times a week. Tumor was removed 10 days after the last injection and allowed for measurement of tumor size and volume.

Sheu M.L., Pan L.Y., Hu H.Y., Su H.L., Sheehan J., Tsou H.K, & Pan H.C. (2022). Potential Therapeutic Effects of Thiazolidinedione on Malignant Glioma. International Journal of Molecular Sciences, 23(21), 13510.

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Molecular Mechanisms of Inflammation Regulation

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The recombinant human IL-1β, PGE₂, forskolin, PGD₂, 15d-PGJ₂ and the inhibitors including NS398, LY294002, SB203580 and SP600125 were obtained from Sigma-Aldrich Corp. Quinazoline (QNZ) was obtained from Enzo Life Sciences (Farmingdale, NY, USA). Rofecoxib was purchased from Tangchao Chemical Inc. (Xian, SX, China). HO-1 siRNA was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies specific for β-actin, COX-2, Akt, p-Akt (Ser 473), p38, p-p38 (Thr 180/Tyr 182), c-Jun, p-c-Jun (Ser 63), NF-κB, p-NF-κB (Ser 536), p-NF-κB (Ser 276), IL-1β, FGF-2, MMP-1, MMP-11 and HO-1 were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Recombinant human FGF-2 was obtained from Raybiotech, Inc. (Norcross, GA, USA). The PGE₂ and IL-1β enzyme immunoassay kits were from Cayman Chemical (Ann Arbor, MI). 15d-PGJ₂ enzyme immunoassay kits were obtained from Assay Designs (Ann Arbor, MI, USA). FGF-2 enzyme immunoassay kits were purchased from Raybiotech, Inc. (Norcross, GA, USA). All reagents for qRT-PCR and SDS-PAGE experiments were purchased from Bio-Rad Laboratories. All other reagents were from Invitrogen (Carlsbad, CA), unless otherwise specified.

Guan P.P., Guo J.W., Yu X., Wang Y., Wang T., Konstantopoulos K., Wang Z.Y, & Wang P. (2015). The Role of Cyclooxygenase-2, Interleukin-1β and Fibroblast Growth Factor-2 in the Activation of Matrix Metalloproteinase-1 in Sheared-Chondrocytes and Articular Cartilage. Scientific Reports, 5, 10412.

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Macrophage Polarization and Nrf2 Activation

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Cells were left untreated (Con) or incubated with 5-ng ml⁻¹ LPS and 10-ng ml⁻¹ IFNγ (M1), 10-ng ml⁻¹ IL-4 (M2), or 5-ng ml⁻¹ LPS (LPS). LPS from Escherichia coli 0111:B4 was purchased from Sigma-Aldrich, and IFNγ and IL-4 were from R&D Systems. For pharmacological Nrf2 activation, cells were treated with 100-μM diethylmaleate (DEM) or 15-deoxy-Δ^12,14-prostaglandin J₂ (15d-PGJ₂). These Nrf2 inducers were stored as the solutions dissolved in DMSO (vehicle). DEM was from Wako chemicals, and 15d-PGJ₂ was from Sigma-Aldrich. For detection of ROS, BMDMs were treated with CellROX Deep red reagent (Thermo) during the last 30 min of 4 h stimulation with or without 5-ng ml⁻¹ LPS concomitant treatment with 100-μM DEM or 1-mM NAC. CellROX signal was detected with flow cytometry analysis using FACS-Caliber (BD Biosciences) and FlowJo software (TOMY Digital Biology). For the evaluation of the mRNA stability, 5-μg ml⁻¹ actinomycin D (Sigma-Aldrich) was added to cell cultures pre-treated with LPS for 3 h without removal of the LPS-containing media. Unless stated, cells were stimulated for 6 h in microarray and qRT-PCR analyses, and for 4 h in ChIP-seq, ChIP-qPCR and immunoblot analyses.

Kobayashi E.H., Suzuki T., Funayama R., Nagashima T., Hayashi M., Sekine H., Tanaka N., Moriguchi T., Motohashi H., Nakayama K, & Yamamoto M. (2016). Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription. Nature Communications, 7, 11624.

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Preparation of 15d-PGJ2-Loaded PLGA Nanoparticles

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The 15d-PGJ2-NP production was prepared according to the protocol established by Alves et al. [11 (link)]. The PLGA Polymer (100 mg) was dissolved into 30 mL acetone with 15d-PGJ2 (100 μg; Sigma-Aldrich, St. Louis, MO, USA), sorbitan monostearate (40 mg), and caprylic/capric acid triglyceride (200 mg) the organic phase. Then, polysorbate 80 (60 mg) and deionized water (30 mL) were added, comprising the aqueous phase. Briefly, after the components in both phases were dissolved, the organic phase was gently added to the aqueous phase, and the suspension was maintained under agitation for 10 min. The solvent acetone (60/40 v/v). was removed under evaporation, and the suspension was concentrated to a volume of 10 mL under low pressure, using a rotary evaporator, to obtain a suspension of 15d-PGJ2 with a final concentration of 10 μg/mL. Residual solvent was removed by evaporation until there were no observed traces of acetone in the preparation and analyzed by HPLC, as previously described (Napimoga et al., 2012) [9 (link)].

Uemura L., Baggio Simeoni R., Bispo Machado Júnior P.A., Gavazzoni Blume G., Kremer Gamba L., Sgarbossa Tonial M., Baggio Simeoni P.R., Stadler Tasca Ribeiro V., Silvestre R., de Carvalho K.A., Napimoga M.H., Cesar Francisco J, & Guarita-Souza L.C. (2022). Autologous Bone Marrow Mononuclear Cells (BMMC)-Associated Anti-Inflammatory Nanoparticles for Cardiac Repair after Myocardial Infarction. Journal of Functional Biomaterials, 13(2), 59.

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Molecular Mechanisms of Antifibrotic Compounds

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Curcumin, Y15, 15d-PGJ2 and PD68235 were obtained from Sigma-Aldrich (St Louis, MO, USA). U0126 was obtained from Cell Signaling Technology (Danvers, MA, USA). Imatinib and fasudil were obtained from Nanjing EnoGene Biotechnology (Nanjing, China). Rapamycin was obtained from Xi'an Helin Biological Engineering (Xi'an, China). All these compounds were dissolved in dimethylsulfoxide (DMSO; Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) for experiments. Recombinant rat PDGF was obtained from Cell Sciences (Canton, MA, USA). Primary antibodies against VEGF, p-PI3K, PI3K, p-AKT and AKT were obtained from Nanjing EnoGene Biotechnology (Nanjing, China). Primary antibodies against α-smooth muscle actin (α-SMA), α(I) procollagen, fibronectin, p-PDGF-βR, PDGF-βR, p-FAK, FAK, GTP-RhoA and total-RhoA were obtained from Epitomics (San Francisco, CA, USA). Primary antibodies against PPAR-γ, p-ERK and ERK were obtained from Cell Signaling Technology. Primary antibodies against HIF-1α, VEGF-R2, p-mTOR, mTOR, p-p70S6K, p70S6K and β-actin were obtained from Bioworld Technology (Nanjing, China).

Zhang F., Zhang Z., Chen L., Kong D., Zhang X., Lu C., Lu Y, & Zheng S. (2014). Curcumin attenuates angiogenesis in liver fibrosis and inhibits angiogenic properties of hepatic stellate cells. Journal of Cellular and Molecular Medicine, 18(7), 1392-1406.

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Evaluating Small Molecule Modulators in Cell Signaling

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The following compounds were used in this study: curcumin, compounds 15d-PGJ2 and PD68235 (Sigma, St. Louis, MO, USA); Etoposide (Selleckchem, Houston, TX, USA); P53 inhibitor PFT-α (Nanjing EnoGene Biotech Co., Ltd, Nanjing, China). All these compounds were dissolved in dimethylsulfoxide (DMSO; Sinopharm Chemical Reagent Co., Ltd, Shanghai, China) for experiments. The primary antibodies were used in this study: P16, P53, cyclin D1, cyclin E1, CDK4, CDK6, PPAR-γ, γH2AX and β-actin (Cell Signaling Technology, Danvers, MA, USA); P21 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); Hmga1 (Abcam Technology, Abcam, Cambridge, UK); α1(I)-procollagen and α-SMA (Epitomics, San Francisco, CA, USA).

Jin H., Lian N., Zhang F., Chen L., Chen Q., Lu C., Bian M., Shao J., Wu L, & Zheng S. (2016). Activation of PPARγ/P53 signaling is required for curcumin to induce hepatic stellate cell senescence. Cell Death & Disease, 7(4), e2189-.

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