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(A-D) Relative mRNA levels of genes involved in (A) fatty acid uptake, (B) fatty acid oxidation, (C) de novo lipogenesis, and (D) Lipid storage/droplet-associated proteins in the livers of fed or 16 h fasted Hmgcs2 ΔLiv mice. (E) WB analysis of lipolysis-associated proteins in the liver lysates from Hmgcs2 ΔLiv mice. (F) Relative mRNA levels of ACSL1 isoforms in the livers of Hmgcs2 ΔLiv mice. (G) WB analysis and quantification of ACSL1 in the liver of Hmgcs2 ΔLiv mice. (H) Oil-red-O staining of primary hepatocytes from Hmgcs2 ΔLiv mice and Hmgcs2 F/F mice loaded with 200 mM BSA-PA in the presence or absence of 5mM Triascin C treatment for 16 h (20X magnification). (I and J) WB analysis of ACSL1 in the hepatic mitochondrial and microsomal fractions from 16 h fasted Hmgcs2 F/F mice. (K-M) Thin layer chromatography (TLC) image showing lipid fractions including Phospholipids (PL), <t>Cholesterol/Diacylglycerol</t> (Chol/DG), free fatty acids (FFA), Triacylglycerol (TAG) and ceramides (CE). (L) Quantified band densities for TAG lipid fractions and (M) Quantified ratio of TAG/PL in the liver of fasted Hmgcs2 F/F mice. All the data is presented as mean ± SEM. p < 0.05 (*) or p < 0.01 (**) or p < 0.001 (***) analyzed by One-way ANOVA (Tukey multiple-comparisons test) and Two-tailed Student t -test.
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infinity tm triglyceride reagent kit  (Millipore)
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(A-D) Relative mRNA levels of genes involved in (A) fatty acid uptake, (B) fatty acid oxidation, (C) de novo lipogenesis, and (D) Lipid storage/droplet-associated proteins in the livers of fed or 16 h fasted Hmgcs2 ΔLiv mice. (E) WB analysis of lipolysis-associated proteins in the liver lysates from Hmgcs2 ΔLiv mice. (F) Relative mRNA levels of ACSL1 isoforms in the livers of Hmgcs2 ΔLiv mice. (G) WB analysis and quantification of ACSL1 in the liver of Hmgcs2 ΔLiv mice. (H) Oil-red-O staining of primary hepatocytes from Hmgcs2 ΔLiv mice and Hmgcs2 F/F mice loaded with 200 mM BSA-PA in the presence or absence of 5mM Triascin C treatment for 16 h (20X magnification). (I and J) WB analysis of ACSL1 in the hepatic mitochondrial and microsomal fractions from 16 h fasted Hmgcs2 F/F mice. (K-M) Thin layer chromatography (TLC) image showing lipid fractions including Phospholipids (PL), <t>Cholesterol/Diacylglycerol</t> (Chol/DG), free fatty acids (FFA), Triacylglycerol (TAG) and ceramides (CE). (L) Quantified band densities for TAG lipid fractions and (M) Quantified ratio of TAG/PL in the liver of fasted Hmgcs2 F/F mice. All the data is presented as mean ± SEM. p < 0.05 (*) or p < 0.01 (**) or p < 0.001 (***) analyzed by One-way ANOVA (Tukey multiple-comparisons test) and Two-tailed Student t -test.
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(A-D) Relative mRNA levels of genes involved in (A) fatty acid uptake, (B) fatty acid oxidation, (C) de novo lipogenesis, and (D) Lipid storage/droplet-associated proteins in the livers of fed or 16 h fasted Hmgcs2 ΔLiv mice. (E) WB analysis of lipolysis-associated proteins in the liver lysates from Hmgcs2 ΔLiv mice. (F) Relative mRNA levels of ACSL1 isoforms in the livers of Hmgcs2 ΔLiv mice. (G) WB analysis and quantification of ACSL1 in the liver of Hmgcs2 ΔLiv mice. (H) Oil-red-O staining of primary hepatocytes from Hmgcs2 ΔLiv mice and Hmgcs2 F/F mice loaded with 200 mM BSA-PA in the presence or absence of 5mM Triascin C treatment for 16 h (20X magnification). (I and J) WB analysis of ACSL1 in the hepatic mitochondrial and microsomal fractions from 16 h fasted Hmgcs2 F/F mice. (K-M) Thin layer chromatography (TLC) image showing lipid fractions including Phospholipids (PL), Cholesterol/Diacylglycerol (Chol/DG), free fatty acids (FFA), Triacylglycerol (TAG) and ceramides (CE). (L) Quantified band densities for TAG lipid fractions and (M) Quantified ratio of TAG/PL in the liver of fasted Hmgcs2 F/F mice. All the data is presented as mean ± SEM. p < 0.05 (*) or p < 0.01 (**) or p < 0.001 (***) analyzed by One-way ANOVA (Tukey multiple-comparisons test) and Two-tailed Student t -test.

Journal: Research Square

Article Title: Hepatic ketogenesis regulates lipid homeostasis via ACSL1-mediated fatty acid partitioning

doi: 10.21203/rs.3.rs-3147009/v1

Figure Lengend Snippet: (A-D) Relative mRNA levels of genes involved in (A) fatty acid uptake, (B) fatty acid oxidation, (C) de novo lipogenesis, and (D) Lipid storage/droplet-associated proteins in the livers of fed or 16 h fasted Hmgcs2 ΔLiv mice. (E) WB analysis of lipolysis-associated proteins in the liver lysates from Hmgcs2 ΔLiv mice. (F) Relative mRNA levels of ACSL1 isoforms in the livers of Hmgcs2 ΔLiv mice. (G) WB analysis and quantification of ACSL1 in the liver of Hmgcs2 ΔLiv mice. (H) Oil-red-O staining of primary hepatocytes from Hmgcs2 ΔLiv mice and Hmgcs2 F/F mice loaded with 200 mM BSA-PA in the presence or absence of 5mM Triascin C treatment for 16 h (20X magnification). (I and J) WB analysis of ACSL1 in the hepatic mitochondrial and microsomal fractions from 16 h fasted Hmgcs2 F/F mice. (K-M) Thin layer chromatography (TLC) image showing lipid fractions including Phospholipids (PL), Cholesterol/Diacylglycerol (Chol/DG), free fatty acids (FFA), Triacylglycerol (TAG) and ceramides (CE). (L) Quantified band densities for TAG lipid fractions and (M) Quantified ratio of TAG/PL in the liver of fasted Hmgcs2 F/F mice. All the data is presented as mean ± SEM. p < 0.05 (*) or p < 0.01 (**) or p < 0.001 (***) analyzed by One-way ANOVA (Tukey multiple-comparisons test) and Two-tailed Student t -test.

Article Snippet: Serum triglycerides and cholesterol levels were measured using colorimetric Infinity Triglyceride and Cholesterol Reagent kits (Thermofisher Scientific, Middletown, VA).

Techniques: Staining, Thin Layer Chromatography, Two Tailed Test