Polyoxyethylene sorbitan monolaurate

Dulbecco’s modified Eagle’s medium (DMEM)/Nutrient Mixture F-12 Ham, APAP, l-glutamine, l-ascorbic acid, anti-GAPDH antibody and Triton^TM X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Collagenase, insulin human recombinant and cycloheximide were purchased from Wako Pure Chemical Industries (Osaka, Japan). Soybean trypsin inhibitor powder, HEPES buffer (1 M) and Dynabeads^® Protein G were purchased from Thermo Fisher Scientific (Gibco^®; Waltham, MA, USA). 2-Mercaptoethanol, dexamethasone, Nonidet^® P-40 and polyoxyethylene sorbitan monolaurate were purchased from Nacalai tesque (Kyoto, Japan). MG132 (Z-Leu-Leu-Leu-H aldehyde) was purchased from Peptide Institute (Osaka, Japan). Proteasome substrates II, III and VI, and Fluorogenic and anti-rabbit CYP3A1 antibodies were purchased from Merck Millipore (Calbiochem^®; Darmstadt, Germany). Penicillin G potassium and streptomycin were purchased from Meiji Seika Pharma (Tokyo, Japan). Anti-mouse CYP3A1, anti-rabbit CHIP, anti-rabbit gp78-2, anti-rabbit Nrf2 and anti-mouse Ub antibodies were purchased from Santacruz Biotechnology (Dallas, Texas, USA). P450-Glo^TM CYP3A assay with luciferin-IPA was purchased from Promega (Fitchburg, WI, USA).

The powder (100 g) made from the leaves of Albizia richardiana was mixed with 500 mL of 70% (v/v) aqueous methanol for 48 h. The leaf extracts were filtrated using a layer of filter paper (No. 2, 125 mm; Advantec, Toyo Roshi Kaisha Ltd., Tokyo, Japan), and the residue mixture was re-extracted with 500 mL of methanol for 24 h then filtrated again. These two residues of leaf powder were mixed and evaporated with a rotary evaporator at 40 °C. The leaf extracts were dissolved in methanol (250 mL) to produce bioassay concentrations of 3, 10, 30, 100, 300, and 1000 mg dry weight (DW) equivalent extract/mL, and assay tests were conducted with the Albizia richardiana crude extracts against the selected species for this study as recommended by Hossen et al. [38 ]. The exact quantity of residues was transferred to a filter paper (No. 2, 28 mm; Toyo Roshi Ltd., Tokyo, Japan) on 28 mm Petri dishes. The Petri dishes containing the methanolic residues were then desiccated in a draft chamber, and 0.6 mL of 0.05% (v/v) aqueous solution of Tween 20 (polyoxyethylene sorbitan monolaurate; Nacalai Tesque, Inc., Kyoto, Japan) (acts as a surface active agent without any harmful effect) was added to each dish. The seeds or germinated seeds of chosen plants were then placed onto the filter paper in the Petri dishes. Only Tween 20 solution was applied for the control experiments.