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5 protocols using f001 1 1

1

Measurement of Cholesterol and Triglycerides

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The levels of total cholesterol and triglycerides (TG) were determined with commercial kits according to the manufacturer’s instructions. The total cholesterol assay kit (A111-1-1) and triglyceride assay kit (F001-1-1) were purchased from Nanjing Jiancheng Bioengineering Institute.
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2

Comprehensive Analysis of Pork Quality Attributes

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Contents of moisture, crude protein, IMF, ash, and inosinic acid in LT were measured by freeze-dryer (ALPHA 2–4 LSC, Christ Martin GmbH), automatic nitrogen analyzer (8400, FOSS, Denmark), fat analyzer (2055 SOXTEC, FOSS, Denmark), box resistance furnace (SX2-4-10N, Yiheng, Shanghai, China), and high performance liquid chromatography (HPLC, LC-20AD, Shimazu, Japan), respectively, as described previously (16 (link)). Briefly, the contents of moisture, crude protein, IMF, and ash were analyzed by the freeze-drying method, Kjeldahl method, Soxhlet extraction, and burning method, respectively.
The sample preparation procedure comprised homogenization of LT (0.1 g) and 0.9% saline (0.9 mL), and 1,500 r/min for 2 min, followed by centrifugation at 3,500 × g for 15 min at 4°C to collect the supernatant. Then, contents of cholesterol, triglyceride, and malondialdehyde (MDA) and activities of total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) were determined according to the instructions (F001-1-1, F002-1-1, A003-2-2, A015-1-2, A005-1-2, A001-1-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The plate was read by a multi-functional enzyme labeling instrument (Spectra Max M5, Molecular Devices, USA) at 532 nm (MDA), 520 nm (T-AOC), 412 nm (GSH-Px), and 450 nm (SOD).
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3

Serum Lipid Profile Determination

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Blood samples were collected and left for 2–3 h at room temperature. The serum was centrifuged at 3000 rpm for 15 min, divided, and stored at −80 °C. TC, TG, LDL-C, and HDL-C levels in serum were determined using commercially available assay kits (cat: F002-1-1, F001-1-1, A113-2-1, and A112-2-1, Nanjing Jiancheng Bioengineering Institute, China).
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4

Lipid Panel Biomarker Measurement

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TC, TG, FFA, LDL-C, and HDL-C levels were measured by corresponding kits according to the manufacturer’s instructions (F001-1-1, F002-1-1, A042-2-1, A112-1-1, and A113-1-1; Jiancheng Bioengineering Institute, Nanjing, China).
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5

Intracellular Lipid and Glucose Quantification

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Intracellular TG and TCH concentrations were measured according to the instructions of the TG (F001-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China) and TCH assay kits (F002-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and the values were corrected using the protein concentration of the samples. The glucose concentration in the medium was measured according to the instructions of the glucose assay kits (A154-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and the values were corrected using the cell numbers of the samples. The cell numbers of the samples were tested by an automated cell counting chamber (SD100-002, Nexcelom Bioscience, Shanghai).
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