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5 protocols using serum triglyceride quantification kit

1

Rodent Metabolic Profile Monitoring

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During the course of the current study, all the rodents underwent weekly body weight and blood glucose estimations. Nonfasting blood glucose level was measured by means of tail blood samples using Glutest mint (Sanwa Kagaku, Aichi, Japan). The rats were euthanized through deep anesthesia at the ages of five, eight, twelve, and sixteen weeks. Subsequently, the chest cavity was opened and blood was collected from the right atrium. The collected blood was centrifuged at 1710 × g for 10 minutes at 4°C, and the serum was collected. Serum triglyceride was measured using a Serum Triglyceride Quantification Kit (Cell Biolabs, San Diego, CA).
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2

Hepatic Metabolite Profiling Protocol

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The hepatic metabolite levels for glycogen, triglyceride, non-esterified free fatty acid (NEFA) and total cholesterol were determined using commercially available kits from Cell Biolabs (CA, USA). Ultrasonic disruption in PBS containing 0.5% Triton X-100 was used to homogenize 200 mg of liver tissue, which was centrifuged at 5000 xg for 10 minutes at 4°C. The supernatant was collected, stored on ice, and used for glycogen and triglyceride quantification using the Glycogen Assay Kit (Cell Biolabs), and Serum Triglyceride Quantification Kit (Cell Biolabs), respectively. For cholesterol and free fatty acid quantification [26 (link)], 10 mg of liver tissue was extracted with 200 μL of a chloroform:isopropanol:NP-40 (7:11:0.1) mixture in a micro-homogenizer. The extract was centrifuged at 15,000 xg for 10 minutes, and the organic phase was transferred to a new tube and air dried at 50ºC to remove the chloroform. The dried lipids were dissolved and homogenized in 200 μL of 1X Assay Diluent via vortexing. Cholesterol and NEFA were quantified using Total Cholesterol Assay Kit (Cell Biolabs) and Free Fatty Acid Assay Kit (Cell Biolabs), respectively.
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Serum Lipid Analysis in Treated Mice

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Blood was collected from treated mice. Then serum was acquired after centrifugation of the blood at 1000 g for 10 min at 4°C, and all the biochemical indexes, including total cholesterol (TCh), triglyceride (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were measured using Total Cholesterol Assay Kit (STA-384, Cell Biolabs), Serum Triglyceride Quantification Kit (STA-396, Cell Biolabs), HDL and LDL/VLDL Cholesterol Assay Kit (STA-391, Cell Biolabs), respectively.18 (link)
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4

Serum and Liver Biomarker Profiling

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For measurement of blood indexes, GTT and ITT were examined as described previously (after 6 h fasting), while other biomedical indexes were measured under fed conditions. Female WT and db/+ mice were sacrificed on GD 20. Samples of blood were centrifuged at 1000 g for 10 min at 4°C to collect serum samples, which were frozen at -80°C for subsequent analyses. Liver tissues were collected and frozen at -80°C for assessments of biochemical indexes. The TCh, LDL, high-density lipoprotein (HDL), MDA and TG levels were evaluated with Total Cholesterol Assay Kit (STA-384, Cell Biolabs), LDL/VLDL and HDL Cholesterol Assay Kit (STA-391, Cell Biolabs), Lipid Peroxidation MDA Assay Kit (S0131, Beyotime, Shanghai, China), and Serum Triglyceride Quantification Kit (STA-396, Cell Biolabs), respectively. Atherogenic index was determined as previously established (29 (link)). Preparation of liver lysates was conducted following previously described protocol (27 (link)). The MDA, GPx, SOD, catalase (CAT), and glutathione (GSH) levels in the liver were determined using Lipid Peroxidation MDA Assay Kit (S0131), Total Glutathione Peroxidase Assay Kit (S0058), Total Superoxide Dismutase Assay Kit with NBT (S0109), Catalase Assay Kit (S0051), and Glutathione Reductase Assay Kit (S0055), obtained from Beyotime Biotechnology (Shanghai, China), respectively.
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5

Triglyceride Quantification in HeLa Cells

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3 × 106 HeLa cells were suspended in 250 µl of PBS containing 1% Triton X-100. The supernatant was subjected to the colorimetric TG quantification assay (Serum Triglyceride Quantification Kit; CELL BIOLABS). The TG density shown in Fig. S2 is that in the supernatant.
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