Responsiveness of PNPLA3 and lipid-related transcription factors is dependent upon fatty acid profile in primary bovine hepatocytes

Figures

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  Figure 1. Primary bovine hepatocytes stained with DAPI (nuclei; blue), Alexa Fluor 488 Phalloidin (F-actin; green), and Cy3 for patatin-like phospholipase domain-containing protein 3 (PNPLA3; red). A common trait in hepatocytes, some hepatocytes display binucleated nuclei and some do not. The cytoskeletal protein F-actin outlines the shape of the cells. Appearance of PNPLA3 in red shows it scattered throughout the cytoplasm of the cells, where lipolytic activity occurs.

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  Figure 2. Gene expression and protein abundance of patatin-like phospholipase domain-containing protein 3 (PNPLA3) in response to fatty acid (FA) mixture representing liver lipid accumulation (ACCUM; open bars and dotted line) and fatty acid mixture representing liver lipid clearing (RECOV; closed bars and dashed line) at different concentrations in primary bovine hepatocytes. Data is presented as least squares mean ± standard error of the means. Quadratic relationships are displayed if P ≤ 0.15 for a main effect.

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  Figure 3. Protein abundance of abhydrolase domain containing 5 (ABHD5), adipose triglyceride lipase (ATGL), hormone sensitive lipase (HSL), phosphorylated HSL (PHSL), perilipin (PLIN), and phosphorylated PLIN (PPLIN) in response to fatty acid mixture representing liver lipid accumulation (ACCUM; open bars and dotted line) and fatty acid mixture representing liver lipid clearing (RECOV; closed bars and dashed line) at different concentrations in primary bovine hepatocytes. Data is presented as least squares mean ± standard error of the means. Quadratic relationships are displayed if P ≤ 0.15 for a main effect.

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  Figure 4. Gene expression of transcription factors forkhead box O1 (FOXO1), liver X receptor α (LXRA), liver X receptor β (LXRB), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1A), peroxisome proliferator-activated receptor α (PPARA), sirtuin 1 (SIRT1), and sirtuin 3 (SIRT3) in response to fatty acid mixture representing liver lipid accumulation (ACCUM; open bars and dotted line) and fatty acid mixture representing liver lipid clearing (RECOV; closed bars and dashed line) at different concentrations in primary bovine hepatocytes. Data is presented as least squares mean ± standard error of the means. Quadratic relationships are displayed if P ≤ 0.15 for a main effect.

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  Figure 5. Gene expression of fatty acid-mediated genes carnitine palmitoyltransferase 1A (CPT1A), carnitine palmitoyltransferase 2 (CPT2), fatty acid synthase (FASN), and uncoupling protein 2 (UCP2) in response to fatty acid mixture representing liver lipid accumulation (ACCUM; open bars and dotted line) and fatty acid mixture representing liver lipid clearing (RECOV; closed bars and dashed line) at different concentrations in primary bovine hepatocytes. Data is presented as least squares mean ± standard error of the means. Quadratic relationships are displayed if P ≤ 0.15 for a main effect.

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  Figure 6. Gene expression and protein abundance of carbohydrate response element binding protein (ChREBP) and sterol regulatory element-binding transcription factor 1c (SREBP1c) in response to fatty acid mixture representing liver lipid accumulation (ACCUM; open bars and dotted line) and fatty acid mixture representing liver lipid clearing (RECOV; closed bars and dashed line) at different concentrations in primary bovine hepatocytes. Data is presented as least squares mean ± standard error of the means. Quadratic relationships are displayed if P ≤ 0.15 for a main effect.

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  Figure 7. Protein abundance of patatin-like phospholipase domain-containing protein 3 (PNPLA3), carbohydrate response element binding protein (ChREBP), and sterol regulatory element-binding transcription factor 1c (SREBP1c) in response to individual fatty acids at the concentration physiologically found circulating in serum in dairy cows at 0.5 mM of circulating liver lipid clearing profile (iRECOV) in primary bovine hepatocytes. C14:0, C16:0, C18:0, C18:1 n-9, C18:2 n-6, and C22:6 n-3 are abbreviated as C14:0, C16:0, C18:0, C18:1, C18:2, and C22:6 in the figure for conciseness. Data is presented as least squares mean ± standard error of the means. Differences between individual fatty acids are indicated as alphabetic superscripts; when superscripts do not share the same letter, differences between the fatty acids were found (P ≤ 0.07).

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  Figure 8. Working hypothesis of lipid related genes and proteins in primary bovine hepatocytes during the liver lipid accumulation phase (left panel) and the liver lipid recovery phase (right panel). The gray box insert within each panel represents the associations between PNPLA3, transcription factors, and genes where potential for regulatory influence exists. Relative abundance between panels are shown visually with line weight and quantity. Arrows between genes and proteins indicate associative relationships based on published research across species and the current data and do not denote directionality of regulation. Created with BioRendeder.com.