Formaldehyde

Potassium chloride (KCl), calcium chloride (CaCl₂), magnesium chloride (MgCl₂), sodium chloride (NaCl), and formaldehyde were purchased from Vetec Química Fina Ltda. (Brazil). Sodium bicarbonate (NaHCO₃) and glucose (C₆H₁₂O₆) were purchased from Dinâmica (Brazil). Sodium monobasic phosphate (NaH₂PO₄), sodium hydroxide (NaOH), and hydrochloric acid (HCl) were purchased from Nuclear (Brazil). These substances, except glucose, NaCl, and NaHCO₃, were diluted in distilled water to obtain each solution, which was maintained under refrigeration.
Carbamylcholine hydrochloride (CCh) was purchased from Merck (USA). Cremophor®, thiobarbituric acid, tetramethoxypropane, perchloric acid, Mayer's hematoxylin, and eosin were acquired from Sigma-Aldrich (Brazil). All substances were diluted in distilled water as needed for each experimental protocol. The carbogen mixture (95% O₂ and 5% CO₂) was obtained from White Martins (Brazil).

Potassium chloride (KCl), calcium chloride (CaCl₂), magnesium chloride (MgCl₂), sodium chloride (NaCl), and formaldehyde were purchased from Vetec Química Fina Ltda. (João Pessoa, Brazil). Sodium bicarbonate (NaHCO₃) and glucose (C₆H₁₂O₆) were purchased from Dinâmica (Brazil). Sodium monobasic phosphate (NaH₂PO₄), sodium hydroxide (NaOH), and hydrochloric acid (HCl) were purchased from Nuclear (Brazil). These substances, except glucose, NaCl, and NaHCO₃, were diluted in distilled water to obtain each solution, which were maintained under refrigeration.
Carbamylcholine hydrochloride (CCh) was purchased from Merck (USA). Cremophor®, thiobarbituric acid, tetramethoxypropane, perchloric acid, Mayer's hematoxylin, and eosin were acquired from Sigma-Aldrich (Brazil). All substances were diluted in distilled water as needed for each experimental protocol. The carbogen mixture (95% O₂ and 5% CO₂) was obtained from White Martins (Brazil).

Four-day-old fungal cultures grown on PDA treated as described above were used for all confocal microscopy observations. Mycelial plugs 6 mm in diameter were cut from cultures grown on PDA plates and promptly placed in wells of a 24-wells plate containing fixative (3.7% formaldehyde [Vetec] in phosphate buffered saline [PBS], pH 7.2) and incubated at room temperature for 2 h. Next, the samples were washed twice with PBS and stained as follows. The mycelial samples were incubated in 10 mg mL^-1 Calcofluor White (Sigma-Aldrich) and 0.1 M Tris-hydrochloride (pH 9.0 [Roth]) for 20-min at room temperature and washed three times with ultra-pure water. Afterward, the exopolysaccharide matrix (EPS) was stained with 12.5 mg mL^-1 fluorescein-conjugated concanavalin A (Molecular Probes) in PBS for 45 min at room temperature. After washing three times with PBS, the mycelial plugs were inverted on a slide with 6 μL of glycerol (20% in PBS) and covered with a coverslip. Confocal analysis was performed under an SP8 Leica inverted microscope. All samples were exposed to the same light intensity and for the same time. A representative photograph of each condition was selected.

The following drugs were used to execute the experimental protocols: Croton oil, hexadecyltrimethylammonium bromide (HTAB), tetramethylbenzidine (TMB), dexamethasone, rutin, gallic acid, ascorbic acid, chlorogenic acid, rosmarinic acid, solanine, DPPH, ammonium phosphate monobasic (all from Sigma, St. Louis, MO, USA), hematoxylin-eosin, acetonitrile, butanol, paraffin, ethanol, and Folin-Ciocalteu (all from Merck, Whitehouse Station, New Jersey, USA), Isoflurane (Baxter, São Paulo, Brazil), sodium acetate, acetone, acetic acid, and formaldehyde (all from Vetec, Rio de Janeiro, Brazil).

The R-(-)-apomorphine, malondialdehyde (MDA), 1,1-diphenyl-2-picryl hydrazyl (DPPH) and acetylcholine (ACh) were obtained from Sigma-Aldrich (São Paulo, Brazil). Phenylephrine (Phe) was purchased from Pfizer (Kalamazoo, USA). Magnesium sulphate (MgSO₄), potassium chloride (KCl), calcium chloride (CaCl₂), sodium chloride (NaCl) and formaldehyde were purchased from Vetec Química Fina Ltda. (São Paulo, Brazil). Glucose (C₆H₁₂O₆) and sodium bicarbonate (NaHCO₃) were purchased from Dinâmica (São Paulo, Brazil). Potassium monobasic phosphate (KH₂PO₄), sodium hydroxide (NaOH) and hydrochloric acid (HCl) were purchased from Nuclear (São Paulo, Brazil). These substances, except the glucose, NaCl and NaHCO₃ were diluted in distilled water to obtain each solution, which were maintained under refrigeration.

Bovine serum albumin, 5,5′-dithiobis (2-nitrobenzoic acid) (DTNB), reduced glutathione (GSH), xylenol orange, K2HPO4, KH2PO4, 1 M Tris, ethylenediaminetetraacetic acid, Tris HCl (all from Sigma, St. Louis, MO, United States), pyrogallol, absolute ethanol, absolute methanol, ferrous ammonium sulfate, trichloroacetic acid, formaldehyde (all from Vetec, Rio de Janeiro, Brazil), and ultra-pure water from a Milli-Q system were used for eluent preparation.

Magnesium sulfate (MgSO₄), potassium chloride (KCl), calcium chloride (CaCl₂), sodium chloride (NaCl) and formaldehyde were purchased from Vetec Química Fina Ltda. (Brazil). Glucose (C₆H₁₂O₆) and sodium bicarbonate (NaHCO₃) were purchased from Dinâmica (Brazil). Potassium monobasic phosphate (KH₂PO₄), sodium hydroxide (NaOH) and hydrochloric acid (HCl) were purchased from Nuclear (Brazil). These substances, except glucose, NaCl and NaHCO₃ were diluted in distilled water to obtain each solution, which were maintained under refrigeration.
Phenylephrine (Phe) was purchased from Pfizer (USA). Acetylcholine (ACh), sodium nitroprusside (SNP), L-N^ω-nitroarginine methyl ester (L-NAME), indomethacin, tempol, apocynin, Cremophor® and R-(-)-apomorphine were obtained from Sigma-Aldrich (Brazil). All substances were diluted in distilled water as needed for each experimental protocol. The carbogen mixture (95% O₂ and 5% CO₂) was acquired from White Martins (Brazil).