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Figure 3 | Molecular Medicine

Figure 3

From: p38 Mitogen-Activated Protein Kinase and Liver X Receptor-α Mediate the Leptin Effect on Sterol Regulatory Element Binding Protein-1c Expression in Hepatic Stellate Cells

Figure 3

LXRα is involved in leptin inhibition of SREBP-1c expression in cultured HSCs. (A, C, D) Transfection assay for analysis of the activity of Sp1, NF-Y or LXR. HSCs were transfected with plasmid pSP1-Luc (A), pNFY-Luc (C) or pLXRE-Luc (D) and treated with leptin at increasing doses for 24 h (n = 6). *P < 0.05 (A, D), #P > 0.05 (C) versus the respective control without leptin. (B, E) Transfection assay for analysis of SREBP-1c promoter activity. HSCs were transfected with a fixed amount of a DNA mixture (including pSREBP1c-Luc, pSP1 or pLXRα, pRL-TK and empty vector) per well and treated with or without 100 ng/mL leptin for 24 h (n = 6). The empty vector was used to ensure the equal amount of total DNA in transfection assay. (P > 0.05 versus the control (the first column on the left) (B). #P > 0.05 versus cells without pSP1 and leptin (the first column on the right) (B). *P < 0.05 versus cells without pLXRα (E). (F) Western blot analysis of LXRα protein level (n = 3). HSCs were incubated with different doses of leptin for 24 h. (G) Transfection assay for analysis of the role of LXR binding sites in leptin inhibition of SREBP-1c promoter activity. HSCs were transfected with pmut-LXRE-Luc or pSREBP1c-Luc and treated with or without 100 ng/mL leptin for 24 h. The luciferase activity in cells without leptin for each construct was expressed as 100% activity after Renilla luciferase activity normalization (n = 6). *P < 0.05 versus cells without leptin for each construct. #P < 0.05 versus cells with pSREBP1c-Luc (leptin treatment).

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