Potassium chloride (kcl)

The following reagents were used: N-methyl-2-pyrollidone (NMP) (Merck, Darmstadt, Germany), isopropyl alcohol (IPA) (special purity, Chimmed, Moscow, Russia), lithium hydroxide (≥99.0%, Sigma Aldrich, St. Louis, MO, USA), hydrochloric acid (special purity, 35–38%, Chimmed, Moscow, Russia), nitric acid (special purity, 60%, Chimmed, Moscow, Russia), hydrogen peroxide special (purity, 30%, Chimmed, Moscow, Russia), potassium chloride (reagent grade, Chimmed, Moscow, Russia), potassium hydroxide (reagent grade, Chimmed, Moscow, Russia), sodium chloride (reagent grade, Chimmed, Moscow, Russia), calcium chloride (reagent grade, Chimmed, Moscow, Russia), sodium bicarbonate (reagent grade, Chimmed, Moscow, Russia), N-acetyl-L-methionine (AM) (99%, J&K Scientific, San Jose, CA, USA), L-carnitine (CT) (>98%, Alfa Aesar, Haverhill, MA, USA), L-lysine (LS) (>99%, Alfa Aesar, Haverhill, MA, USA), deionized water (resistance 18.2 MOhm).
The membranes used were Nafion^® 212 (The Chemours Company, Wilmington, DE, USA) and Aquivion E87-05S (Solway, Brussels, Belgium). Nafion^® 212 film (The Chemours Company, Wilmington, DE, USA) and Aquivion PW79S powder (Solway, Brussels, Belgium) were used to obtain PFSA polymer dispersions. The main characteristics of the materials used are shown in Table 1.

Cells were seeded in 60 mm Petri dishes (SPL Lifescience, Pochon, Republic of Korea) at 3 × 10⁵ per dish and incubated overnight under standard conditions. Afterward, the cells were treated with CBL0137 at concentrations of 0.6 μM and 1.2 μM or DMSO (0.1%) as vehicle control. After 24 h, the cells were washed with PBS and lysed in buffer for nuclear extraction and fractionation (Abcam protocol: 20 mM HEPES pH 7.4 (PanEco, Moscow, Russia), 10 mM KCl (CHIMMED, Moscow, Russia), 2 mM MgCl₂ (CHIMMED, Moscow, Russia), 1 mM EDTA (PanEco, Moscow, Russia), 1 mM EGTA (AppliChem, Darmstadt, Germany), 20 mM dithiothreitol (SERVA, Heidelberg, Germany), and Complete Protease Inhibitor Cocktail (Roche, Basel, Switzerland)). Briefly, the cells were lysed for 15 min on ice, and then cell suspension was performed through a 26- and 23-gauge needle with the removal of supernatant (for the mechanical destruction of the cytoplasmic and nuclear membrane, respectively). The precipitate was resuspended in deionized water with Complete Protease Inhibitor Cocktail (Roche, Basel, Switzerland) and then homogenized with Soniprep 150 Plus ultrasonic homogenizer (MSE, London, UK) for 3 s at a power of 2A on ice. The nuclear fraction was used to analyze the enzymatic activity of HATs as well as the level of HDAC1 and HDAC3 protein expression.

Ultrapure deionized water (Milli-Q^®, 18 MΩ cm, Millipore, Burlington, MA, USA) was used to prepare all solutions. All chemicals used were of at least analytical grade. Inorganic compounds (NaOH, HCl, KCl, KH₂PO₄, K₂HPO₄) were obtained from Chimmed (Russia). Folin and Ciocalteu’s phenol reagent, 2,2′-azobis (2-amidino-propane) dihydrochloride (AAPH), fluorescein, 6-hydroxy-2,5,7,8-tetra-methylchroman-2-carboxylic acid (Trolox), hydroxybenzoic acid (HBA), vanillic acid, syringic acid, gallic acid, fumaric acid, ferulic acid, sinapic acid, ascorbic acid, and vitamin E were procured from Sigma-Aldrich (Burlington, MA, USA). Standard samples of HS, including Suwannee River humic acid (SRHA) and fulvic acid (SRFA) and dissolved organic matter (SRDOM), were kindly provided by I.V. Perminova, coordinator of the Russian chapter of the International Humic Substances Society (IHSS, St. Paul, MN, USA). Commercially available coal humic materials CHA-AGK and CHA-ALD were purchased from Biotechnology Ltd. (Moscow, Russia) and Sigma-Aldrich (Burlington, MA, USA), respectively.

Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), N-methyl-D-aspartate (NMDA), Bicuculline methochloride, L-NAME hydrochloride, Glycine, Domoic acid, (R)-(+)-Methanandamide, L-arginine, Acetyl-L-carnitine chloride, UK 14304, Telenzepine dihydrochloride (Tocris Bioscience, UK); Hank’s Balanced salt solution (HBSS), neurobasal medium, B-27 Supplement, fetal bovine serum, fetal calf serum (Gibco); penicillin-streptomycin solution (Dalhimfarm, Russia); 0.1% poly-l-lysine; L-glutamine, L-glutamate, NBQX hydrate, (+)-MK-801 hydrogen maleate, NAAG, Betaine monohydrate, L-carnitine hydrochloride, P-F-HHSiD (p-Fluorohexahydro-sila-difenidol hydrochloride), Methoctramine hydrate, Nystatin (Sigma-Aldrich, USA); KCl (Chimmed, Russia); embryo calf serum (MP Biomedicals, USA); 4% gentamicin (Dalhimfarm, Russia); versene (Paneco, Russia); Fura-2 AM (Invitrogen, USA).
All animal studies were performed in accordance with the legal requirements and were approved by the Animal Ethic Committees of both institutes.