Human umbilical vein endothelial cells (HUVEC) were purchased from PromoCell (Heidelberg, Germany) and grown in endothelial cell growth medium (PromoCell) supplemented with gentamycin (50 µg mL−1) and amphotericin B (50 ng mL−1). For all experiments, we used HUVECs pooled from up to four donors, which did not exceed passage 4. Human embryonic kidney (HEK) 293 cells (ATCC CRL-1573) were grown in DMEM medium (Invitrogen, Cergy Pontoise, France) supplemented with 10% fetal calf serum (FCS), 100 IU mL−1 penicillin, and 100 µg mL−1 streptomycin (Invitrogen, Cergy Pontoise, France). C166 mouse endothelial cells (accession number CRL-2581) and LLC1 mouse lung cancer cells (accession number CRL-1642) were grown in DMEM medium (Invitrogen, Cergy Pontoise, France). Media were supplemented with 10% fetal calf serum (FCS), 100 IU mL−1 penicillin and 100 µg mL−1 streptomycin. As positive control for apoptosis assays, LLC1 mouse lung cancer cells were treated with 100 nmol/L Staurosporine (Sigma, St. Louis, MO, USA) overnight. For RNA isolation and quantitative RT-PCR experiments, HUVEC and LLC1 cells were maintained for 48 h (HUVEC) or 24 h (LLC1) in medium in the presence of GW0742 (Selleckchem, Houston, TX, USA) or GSK3787 (Selleckchem) dissolved in dimethyl sulfoxide (DMSO) at concentrations of 1 µmol/L. Controls were treated with vehicle (0.1% DMSO) only [6 (link),16 (link)].
Gw0742
Manufactured by Selleck Chemicals
Sourced in United States
GW0742 is a synthetic compound used in laboratory research. It functions as a selective agonist for the peroxisome proliferator-activated receptor delta (PPARδ) nuclear receptor. The core function of GW0742 is to activate PPARδ, which plays a role in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Gsk3787, by Selleck Chemicals (1 mentions)
Human umbilical vein endothelial cells (huvec), by Selleck Chemicals (1 mentions)
Staurosporine, by Merck Group (1 mentions)
Human umbilical vein endothelial cells (huvec), by PromoCell (1 mentions)
Endothelial cell growth medium, by PromoCell (1 mentions)
Gw3965, by Selleck Chemicals (1 mentions)
Opti mem media, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Nc sirna, by GenePharma (1 mentions)
Inositol, by Merck Group (1 mentions)
Pioglitazone, by Selleck Chemicals (1 mentions)
5 protocols using gw0742
1
HUVEC Culture and Treatment Protocols
2
Gingival Response to LXR and PPAR Agonists
3
PPAR delta modulation in liver cells
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For PPAR δ activation, we use 0.5 μmol/L GW0742 (Selleck Chemicals, Houston, TX, USA), a PPAR δ-selective agonist, for 24h. For PPAR δ inhibition, the siRNA target sequence used for PPAR δ knockdown (si-PPAR δ). The PPAR δ-specific siRNA gene sequence was 5'-GCAAACCCUUCAGUGAUAUTT -3' and the gene sequence of negative control siRNA (N.C. siRNA) was 5'-UUCUCCGAACGUGUCACGUTT -3'. PPAR δ siRNA and N.C. siRNA was purchased from GenePharma (GenePharma, Shanghai, China). L02 cells were used transfected with 20 pmol of N.C. siRNA or PPAR δ-specific siRNA, using Lipofectamine 2000 and Opti-MEM media (Life Technologies, Gaithersburg, MD, USA) after seeding cells attached to the bottom. After 24h PPAR δ activation or transfection, L02 cells were harvested and immediately subjected to isolation of total RNA and protein. For confirming the PPAR δ knockdown and over expression efficiency, real-time-PCR and Western blot analyses were performed on the isolated total RNA and protein, respectively. Negative control siRNA (N.C. siRNA) or PPAR δ siRNA-transfected L02 cells were incubated in 0.4mM palmitic acid or/and 800ng/ml LPS for 24h. L02 cells were exposed to 0.4mM palmitic acid (PA) or (and) 800ng/ml LPS with or without 1nM GW0742 treatment.
4
Tumor Growth Regulation via PPARβ/δ
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All animal work was conducted according to national and international guidelines and was approved by the local ethics committee (Comité Institutionnel d‘Éthique Pour l‘Animal de Laboratoire Azur (Ciepal), agreement number: PEA-NCE/2013/334). PPARβ/δ-flox+/− [12 (link)] and Tie2-CreERT2 [13 (link)] animals were crossed to generate Tie2-CreERT2;PPARβ/δ-flox+/− mice, further referred to as Tie2-CreERT2;PPARβ/δ. The Tie2-CreERT2-line was back-crossed four times onto C57BL/6J. Age- and sex-matched Tie2-CreERT2;PPARβ/δ animals were injected for one week intraperitoneally either with sunflower oil (vehicle) or Tamoxifen dissolved in sunflower oil in a dose of 33 mg/kg per day [10 (link),14 (link),15 (link)]. Tie2-CreERT2 animals injected with Tamoxifen served as additional controls. One week after the last Tamoxifen or vehicle treatment, 1 × 106 LLC1 tumor cells were injected subcutaneously. Tumors and organs were collected after three weeks. For treatment with the PPARβ/δ agonist, ten-week-old male C57BL/6J (Janvier, France) mice were subcutaneously injected with 1 × 106 LLC1 tumor cells. GW0742 (Selleckchem, Houston, TX, USA) dissolved in DMSO was then subcutaneously injected at 1 mg/kg once every second day (100 µL). Controls received 100 µL DMSO injections [8 (link)].
5
Inositol and Serotonin Signaling Assay
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Inositol (catalogue number PHR1351) was acquired from Sigma (St. Louis, MO, USA). The anti-serotonin antibody was purchased from Abcam (catalog number ab66047). RS-127,445 (item number R2533) and serotonin powder (item number H9523) were both obtained from Merck (Kenilworth, NJ, USA). GW0742 (item number S8020) and Pioglitazone (item number AD-4833) along with asenapine maleate (item number S1283) and myo-Inositol (item number S4530) were purchased from Selleckchem (Houston, TX, USA). A DMSO solution was used to dissolve all the chemicals.
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