Gw6471

OEA and PEA were purchased from Tocris. OEA and PEA were dissolved in ethanol until use, and they were used for cultures after further dilution at 10% ethanol in Neurobasal, at doses of 0, 5, 10, 20, and 40 μM. The selective PPARα antagonist GW6471 or [(2S)-2-[[(1Z)-1-Methyl-3-oxo-3-[4-(trifluoromethyl)phenyl]-1-propenyl] amino]-3-[4-[2-(5-methyl-2-phenyl-4-oxazolyl)ethoxyphenylpropyl-carbamic acid ethyl ester was purchased from Tocris, and it was dissolved in 10% dimethyl sulfoxide (DMSO) until use (IC₅₀ value of GW6471 is 0.24 μM). The selective antagonist of vanilloid TRPV1 SB 452533 or N-(2-Bromophenyl)-N’-[2-[ethyl(3-methylphenyl)amino]ethyl]-urea was purchased from Tocris, and it was dissolved in 10% DMSO until use (pIC₅₀=7.0). The selective antagonist of vanilloid TRPV4, RN1734, or 2,4-Dichloro-N-isopropyl-N-(2-isopropylaminoethyl)benzene sulfonamide, was purchased from Tocris (IC₅₀ value of RN1734 is 3.2 μM for rTRPV4). Finally, GSK1016790A, selective agonist of vanilloid TRPV4, was purchased from Sigma-Aldrich. GSK1016790A or (N-((1S)-1-{[4-((2S)-2-{[(2,4-Dichlorophenyl)sulfonyl]amino}-3-hydroxypropanoyl)-1-piperazinyl]carbonyl}-3-methylbutyl)-1-benzothiophene-2-carboxamide) is known to evoke a dose-dependent activation of TRPV4 whole-cell currents at concentrations above 1 nM.

To study the role of PPAR-α inhibition, PPAR-α-mediated effects of high AEA levels in FAAH^−/− mice were blocked with selective PPAR-α antagonist GW6471 (Tocris Bioscience, Bristol, UK). To show PPAR-α specificity for FAAH^−/− animals, we treated WT mice with GW6471 and did not observe any changes compared to WT, indicating FAAH^−/− specificity of GW6471 treatment in the presented study. Proof of principle was executed by blocking the effect of PPAR-α agonist WY14.634 (0.3 mg/kg in 10% DMSO with 0.9% saline, i.v. injection in the jugular vein 30 min before I/R) [69 (link)] on PPAR-α downstream gene upregulation with antagonist GW6471 in WT as previously described in two different in vivo studies in mice [66 (link),70 (link)]. Intravenous injection of 100µL of either GW6471 (1 mg/kg i.v.) or its vehicle (10% DMSO with 0.9% saline) was performed daily 30 min before LAD occlusion.

AM580 (Tocris, #0760, Germany), ER50891 (Tocris, #3823, Germany), CD2314 (Tocris, #3824, Germany), LE135 (Tocris, #2021, Germany), CD437 (Sigma, #C5865, USA), MM11253 (Tocris, #3822, Germany), ATRA (Sigma, #R2625, USA), BMS493 (Sigma, #B6688, USA), GW7647 (Tocris, #1677, Germany), GW6471 (Tocris, #4618, Germany), GW0742 (Tocris, #2229, Germany), GSK3787 (Tocris, #3961, Germany), Troglitazone (Sigma, #T2573, USA), T0070907 (Sigma, #T8703, USA), T0901317 (Sigma, #T2320, USA), SR9238 (Tocris, #5854, Germany), GC1 (Tocris, #4554, Germany), Calcifediol (Tocris, #4036, Germany), SR9011 (Sigma, #SML2067, USA), SR8278 (Tocris, #4463, Germany), CD3254 (Tocris, #3302, Germany), HX531 (Sigma, #SML2170, USA), Testosterone (Aladdin, #T102169, China), Nilutamide (Sigma, #N8534, USA), XCT790 (Sigma, #X4753, USA), Dexamethasone (Sigma, #D4902, USA), Mifepristone (Sigma, #M8046, USA), Corticosterone (Aladdin, #C104537, China), Eplerenone (Sigma, #E6657, USA), Doxorubicin (Solarbio, #D8740, China), Blasticidin (Beyotime, #ST018, China), Puromycin (Beyotime, # ST551, China).