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3 protocols using linolenic acid

1

Fatty Acid Analysis with Spectroscopic Techniques

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Linolenic acid was from Wako Pure Chemical Industries, Ltd. (Tokyo, Japan). Linoleic acid, γ-linolenic and arachidonic acid were obtained from Sigma Aldrich Co. (St. Louis, MO). Oleic acid and α-(4-pyridyl-1-oxide)-N-tert-butylnitrone (4-POBN) were purchased from Tokyo Kasei Kogyo, Ltd. (Tokyo, Japan). Water used in these experiments was purified by passing through AUTOPURE WT101UV (Nihon Millipore Kogyo K.K., Yonezawa, Japan) after distillation. All other chemicals used were of analytical grade.
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2

Fatty Acid Salt Formulations for Biological Applications

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Sodium oleate, potassium oleate, sodium laurate, and sodium linoleate (shown in Fig. 9) were obtained from TCI (Tokyo, Japan). Linolenic acid, sodium carbonate, sodium bicarbonate (anhydrous), 8 M sodium hydroxide solution, D( +)-glucose, sucrose, and calcium chloride (anhydrous) were purchased from the FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). We obtained DAIGO soybean-casein digest broth from the NIHON Pharmaceutical Co., Ltd. (Tokyo, Japan) and Bacto yeast extract from BD Japan (Tokyo, Japan). Crystal violet, 99.5% ethanol, and glutaraldehyde solution were purchased from Sigma-Aldrich (St. Louis, MO, USA), while PBS solution was obtained from the Nippon Gene Co., Ltd. (Tokyo, Japan). Polymethyl methacrylate (PMMA) prepared by the KYOYUKAI Co., Ltd. (Tokyo, Japan) was cut into 1 cm × 1 cm × 2 mm pieces and used as the substrate.

Structural formulas of fatty acid salts used in this study: (a) potassium oleate, (b) sodium linoleate, (c) sodium linolenate, and (d) sodium laurate.

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3

Soybean Larvae Metabolite Analysis

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Plants and Insects Soybean (Var. Enrei) and S. litura larvae were reared as previously described (Murakami et al. 2014) .
Chemicals L-α-[ 13 C 9 , 15 N]Phenylalanine was purchased from Cambridge Isotope Laboratories, Inc. (Tewksbury, MA, USA). Daidzein and formononetin were purchased from LKT laboratories, Inc. (St. Paul, MN, USA). Genistein and apigenin were purchased from Tokyo Chemical Industry Co, Ltd. (Tokyo, Japan). Kaempferol and 4′,7-dihydroxyflavone were purchased from Extrasynthese (Genay, France). 7-Hydroxyflavone, daidzin, linolenic acid, linoleic acid, L-glutamine, trimethylamine, and ethyl chloroformate were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Tetrahydrofuran was purchased from Kanto Chemical Co., Inc. (Tokyo, Japan). Gut extracts and elicitor solutions were prepared in phosphate buffer (50 mM Na 2 HPO 4 -NaH 2 PO 4, pH 8).
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