Octanol

MP, EP, PP, BP, BPh, 4-MBC, DEET, E1, E2, octanol, decane, and urea were obtained from Sigma-Aldrich (Germany). Carbon tetrachloride was purchased from Merck (Germany). Chloroform, toluene, cyclohexane, sodium hydrogen phosphate (V), magnesium sulfate (VI) anhydrous, sodium bicarbonate, anhydrous potassium carbonate, sodium nitrate (V), and methanol were obtained from POCH (Poland). Acetic anhydride, ammonium bicarbonate (IV), magnesium chloride, potassium hydrogen phosphate (V), potassium carbonate, and calcium chloride anhydrous were obtained from Chempur (Poland). Stock solutions of each analyte (at 1 mg mL⁻¹ each) were prepared separately in methanol and stored at −18 °C not longer than 1 month. Working solutions were prepared by diluting the stock standard solution in methanol and stored at −18 °C not longer than 2 weeks. Deionized water was from purification system (Milli-Q RG, Millipore, USA) and was stored in glass bottle.

Octanol, heptanol, carbenoxolone, oleamide, LaCl₃, Etd⁺, procion orange MX2R, brilliant blue G (BBG), Evans blue, propidium iodide, 4-Bromo calcium ionophore A23187 were purchased from Sigma-Aldrich (St. Louis, Missouri, USA). The 9-amino-6-Chloro-2-Methoxyacridine (ACMA), DAPI⁺², hexidium iodide, nuclear yellow, BOBO 1-iodide, 7-aminoactinomycin D (7-ADD), YOYO 1-iodide, TOTO 1-iodide and ethidium homodimer–2–bromide obtained from Invitrogen (Massachusetts, USA), indocyanine green from Pulsion medical systems, (Feldkirchen, Germany) methylene blue from Euromed (Santiago, Chile).

Carvedilol, metoprolol, sodium chloride, potassium phosphate monobasic, and sodium phosphate dibasic, hexadecane, octanol, and trifluoroacetic acid (TFA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Water and acetonitrile (Merck KGaA, Darmstadt, Germany) were ultra-performance liquid chromatography (UPLC) purity grade. All other substances were of analytical reagent grade.

MWCNTs (OD: 7–15 nm, Length: 0.5–10 μm), Fe(acac)₃ (>99.9% purity), octanol (99.9%) and octylamine (99.9%) were purchased from Sigma–Aldrich. Sodium dodecyl sulfate (SDS) was purchased from Sinopharm Chemical Reagent Co., Ltd. (China). All other reagents were of analytical grade and used as received.

Ion currents evoked by the puff application of TRPA1 agonists were studied in conventional whole-cell patch-clamp recordings using pCLAMP and Clampfit software. Patch pipettes with a resistance of 5–12 MΩ as measured in the bath were used. The intracellular solution contained (in mM): KCl (12.6), KGlu (131.4), MgCl₂ (2), EGTA (0.5), K₂HPO₄ (8), KH₂PO₄ (2), Mg₂-ATP (2) and Na₄-GTP (0.2). The L-15 medium in the bath was supplemented with 100 μM of FFA (Sigma-Aldrich, catalog # F9005) or 1 mM Octanol (Sigma-Aldrich). Voltage ramps from −90 mV to +90 mV over 300 ms were applied to study the current-voltage relationships before, during, and 5 min after (washout) puff stimulation with CA. Each type of supporting cell was held close to its resting potential. In the presence of FFA, these resting potentials were found to be (in mV): Hensen’s cells (0 to +10), Deiters’ cells (−10 to −20), outer pillar cells (−20 to −30), and inner pillar cells (−40 to −50).

Phosphocholine (PC) lipids including DOPC (dioleoyl-PC, or di-18:1-PC), DPPC (dipalmitoyl-PC or di-16:0-PC), POPC (palmitoyl-oleoyl-PC or 16:0-18:1-PC), di16:1-Δ9cis-PC, di18:1-Δ6cis-PC, di14:1-Δ9cis-PC, lyso(18:0)-PC, and palmitoyl sphingomyelin (PSM or 18:1-16:0 SM) were from Avanti Polar Lipids (Alabaster, AL). Texas Red dihexadecanoyl-phosphoethanolamine was from Life Technologies (Grand Island, NY), and cholesterol was from Sigma Aldrich (St. Louis, MO). Lipid structures appear in Fig. S1. Stock solutions of laurdan (Invitrogen, Carlsbad, CA) and C-laurdan (a gift from B. R. Cho, Seoul, Korea) were prepared in ethanol and dimethyl sulfoxide (DMSO). Alcohols (ethanol, propanol, butanol, pentanol, hexanol, octanol, decanol, tetradecanol, hexadecanol, propofol, and 2,6-di-tert-butylphenol), DMSO, and all additional reagents were from Sigma Aldrich unless specified. All alcohols were purchased at their highest available purity. All materials were used as from the manufacturer without further purification.