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Regulation of the angiopoietin-like protein 3 gene by LXR

Kaplan, Rebecca, Zhang, Theresa, Hernandez, Melba, Gan, Frank Xiaodong, Wright, Samuel D., Waters, M.Gerard, Cai, Tian-Quan·2003·Journal of Lipid Research
DOI 10.1194/jlr.m200367-jlr200
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Fig. 1 Cholesterol feeding raises the level of Angptl3 mRNA in mouse liver. Mice were fed with a normal chow diet or with the same supplemented with 4% of cholesterol for 7 days. On the 8th day, mice were euthanized, RNAs were prepared from the liver samples, and the relative levels of Angptl3 and CYP7A mRNAs were measured as described in Materials and Methods. Results are normalized to the values obtained from normal chow diet fed mice, and data are shown as the means ± SE of each group (n = 5).
Fig. 1 Cholesterol feeding raises the level of Angptl3 mRNA in mouse liver. Mice were fed with a normal chow diet or with the same supplemented with 4% of cholesterol for 7 days. On the 8th day, mice were euthanized, RNAs were prepared from the liver samples, and the relative levels of Angptl3 and CYP7A mRNAs were measured as described in Materials and Methods. Results are normalized to the values obtained from normal chow diet fed mice, and data are shown as the means ± SE of each group (n = 5).CC-BY · From the PDF
Fig. 2 LXR-selective agonist T0901317 increases levels of plasma lipids and Angptl3 mRNA in the liver. Mice were orally treated with either vehicle or 10 mpk (mg compound/kg of animal body weight/day) of T0901317 for 7 days. On the 8th day, mice were euthanized, and (A) plasma levels of total cholesterol (TC), HDL cholesterol (HDL-C), and triglycerides (TG), or (B) levels of Angptl3 mRNA in the liver were measured. Lipid data are reported as the percent of changes from control mice (A, n = 10); Relative Angptl3 mRNA levels in treated mice are calculated by normalizing to the values obtained from control mice (B, n = 5), and data are shown as the mean ± SE of each group.
Fig. 2 LXR-selective agonist T0901317 increases levels of plasma lipids and Angptl3 mRNA in the liver. Mice were orally treated with either vehicle or 10 mpk (mg compound/kg of animal body weight/day) of T0901317 for 7 days. On the 8th day, mice were euthanized, and (A) plasma levels of total cholesterol (TC), HDL cholesterol (HDL-C), and triglycerides (TG), or (B) levels of Angptl3 mRNA in the liver were measured. Lipid data are reported as the percent of changes from control mice (A, n = 10); Relative Angptl3 mRNA levels in treated mice are calculated by normalizing to the values obtained from control mice (B, n = 5), and data are shown as the mean ± SE of each group.CC-BY · From the PDF
Fig. 3 Concentration-dependent induction of Angptl3 expression by LXR agonists. HepG2 cells (5 × 10 5 cells/well) were plated in 6-well plates and treated with (A) buffer, increasing concentrations of 22( R )-OH Ch or 30 𝛍 M 22( S )-OH Ch or (B) buffer, or increasing concentrations of T0901317. Following H11601 16 h of incubation at 37 ° C, RNA samples were prepared from the cultured cells, and expression levels of Angptl3 mRNA were measured as described in Materials and Methods. Results are normalized to the values obtained from control cells, and data are shown as the means of duplicate determinations.
Fig. 3 Concentration-dependent induction of Angptl3 expression by LXR agonists. HepG2 cells (5 × 10 5 cells/well) were plated in 6-well plates and treated with (A) buffer, increasing concentrations of 22( R )-OH Ch or 30 𝛍 M 22( S )-OH Ch or (B) buffer, or increasing concentrations of T0901317. Following H11601 16 h of incubation at 37 ° C, RNA samples were prepared from the cultured cells, and expression levels of Angptl3 mRNA were measured as described in Materials and Methods. Results are normalized to the values obtained from control cells, and data are shown as the means of duplicate determinations.CC-BY · From the PDF
Fig. 4 Comparison of the genomic regions upstream of Angptl3 in human and mouse. Genomic regions spanning Angptl3 in human and mouse were compared using mVISTA (http://www.gsd.lbl.gov/vista). Conserved regions with an identity greater than or equal to 50% between human and mouse sequences are shown as peaks of similarity (vertical axes) relative to the positions along the mouse genomic sequence (horizontal axes). Peaks representing highly conserved regions (identity > 75% over > 100 bp) are shaded if they overlap with non-coding regions, or marked by slanted lines if they overlap with exons. There are two highly conserved non-coding regions upstream of human and mouse Angptl3: the 500 bp region immediately upstream and the 150 bp region 2.5 kb upstream of the gene.
Fig. 4 Comparison of the genomic regions upstream of Angptl3 in human and mouse. Genomic regions spanning Angptl3 in human and mouse were compared using mVISTA (http://www.gsd.lbl.gov/vista). Conserved regions with an identity greater than or equal to 50% between human and mouse sequences are shown as peaks of similarity (vertical axes) relative to the positions along the mouse genomic sequence (horizontal axes). Peaks representing highly conserved regions (identity > 75% over > 100 bp) are shaded if they overlap with non-coding regions, or marked by slanted lines if they overlap with exons. There are two highly conserved non-coding regions upstream of human and mouse Angptl3: the 500 bp region immediately upstream and the 150 bp region 2.5 kb upstream of the gene.CC-BY · From the PDF
Fig. 5 Analysis of the promoter region of human and mouse Angptl3. The sequence segment overlapping the 5 ′ RACE product is shown in italic, and its 5 ′ end is marked by arrow as the putative transcription start sites (i.e., position + 1 in the sequences). The translational start codon (i.e., ATG) is also marked by an arrow. A number of putative binding sites of transcription factors are predicted in this region. Those conserved between human and mouse are boxed, and those found in only one species are underlined.
Fig. 5 Analysis of the promoter region of human and mouse Angptl3. The sequence segment overlapping the 5 ′ RACE product is shown in italic, and its 5 ′ end is marked by arrow as the putative transcription start sites (i.e., position + 1 in the sequences). The translational start codon (i.e., ATG) is also marked by an arrow. A number of putative binding sites of transcription factors are predicted in this region. Those conserved between human and mouse are boxed, and those found in only one species are underlined.CC-BY · From the PDF
Fig. 6 Effect of LXR agonists on activity of a truncated Angptl3 promoter in HepG2 cells. A fragment of the human Angptl3 promoter (from -953 to + 49 bp) with or without mutations at the DR4 element was linked to the firefly luciferase reporter gene. The resulting plasmid was cotransfected with a control reporter plasmid expressing Renilla luciferase into HepG2 cells. Cells were treated with (A) buffer, 10 𝛍 M of 22( R )-OH Ch, 25-OH Ch, 22( S )-OH Ch, 9-cis-retinoic acid (9-RA), or both 9-RA and 22( R )-OH Ch; or (B) buffer or increasing concentrations of T0901317 or combination of T0901317 (1 𝛍 M) and 9-RA (10 𝛍 M). After 42 h, the cells were lysed and relative luciferase activities were measured. Results are expressed as a ratio between the firefly and Renilla luciferase activities, and data are shown as the means ± SD of triplicates of a representative experiment. Three to four independent experiments were performed with similar results.
Fig. 6 Effect of LXR agonists on activity of a truncated Angptl3 promoter in HepG2 cells. A fragment of the human Angptl3 promoter (from -953 to + 49 bp) with or without mutations at the DR4 element was linked to the firefly luciferase reporter gene. The resulting plasmid was cotransfected with a control reporter plasmid expressing Renilla luciferase into HepG2 cells. Cells were treated with (A) buffer, 10 𝛍 M of 22( R )-OH Ch, 25-OH Ch, 22( S )-OH Ch, 9-cis-retinoic acid (9-RA), or both 9-RA and 22( R )-OH Ch; or (B) buffer or increasing concentrations of T0901317 or combination of T0901317 (1 𝛍 M) and 9-RA (10 𝛍 M). After 42 h, the cells were lysed and relative luciferase activities were measured. Results are expressed as a ratio between the firefly and Renilla luciferase activities, and data are shown as the means ± SD of triplicates of a representative experiment. Three to four independent experiments were performed with similar results.CC-BY · From the PDF

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Citations

Cites

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Metadata

Authors
Kaplan, R., Zhang, T., Hernandez, M., Gan, F. X., Wright, S. D., Waters, M. G., & Cai, T.-Q.
Venue
Journal of Lipid Research
Publisher
Elsevier BV
Published
2003-01
Volume
44
Issue
1
Pages
136-143
ISSN
0022-2275
DOI
10.1194/jlr.m200367-jlr200
Cited by
106
References
2
Language
en
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