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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: Hepatology. 2016 Aug 23;64(5):1559–1576. doi: 10.1002/hep.28747

Figure 1. Loss of NgBR induces lipid accumulation in the mouse liver and hypertriglyceridemia.

Figure 1

(A) NgBR gene expression is decreased in human fatty liver, which is associated with increased expression of lipogenic genes SREBP-1c and FASN. A human liver cDNA panel including 8 healthy control samples and 4 fatty liver samples was purchased from Origene. Gene expression was determined by real-time PCR. * P<0.05; (B-F) NgBR liver-specific knockout (NgBR LivKO) mice were generated as described in the Methods. Both NgBR LivKO and littermate control (NgBR-floxed, no cre) mice were used to conduct the following experiments: (B) PCR analysis of the NgBR gene from mice that are wild-type (NgBR+/+:Creo/o), albumin-Cre (NgBR+/+:Cre+/o or NgBR+/+:Cre+/+), flox/+ (NgBRfl/+:Creo/o), flox/+:albumin-Cre (NgBRfl/+:Cre+/o or NgBRfl/+:Cre+/+), flox/flox (NgBRfl/fl:Creo/o), and flox/flox:albumin-cre (NgBRfl/fl:Cre+/o or NgBRfl/fl: Cre+/+). (C) Expression of NgBR protein and mRNA in the livers of littermate control and NgBR LivKO mice were determined by western blotting with whole liver protein extract (upper panel) or extract of hepatic cells after endothelial cells were removed (bottom panel) and real-time RT-PCR (upper panel) with total RNA isolated from whole liver tissues. *: P<0.05 vs. control (n=4). (D) Littermate control (NgBR-floxed, no cre) and NgBR LivKO female mice at 8 weeks old were fed a Western diet for 4 weeks. NgBR liver-specific knockout increased lipid accumulation determined by Oil Red O staining performed on the frozen liver sections. (E) Quantitative analysis showed that NgBR liver-specific knockout increased the levels of free fatty acid (FFA) and triglyceride (TG). Total lipid extract was prepared from liver tissues of littermate control and NgBR LivKO mice, *: P<0.05 vs. littermate control (n=5). (F) NgBR liver-specific knockout increased the levels of FFA and TG in plasma. Plasma samples were collected from littermate control (NgBR-floxed, no cre) and NgBR LivKO mice. FFA and TG levels were determined by assay kits. *: P<0.05 vs. littermate control (n=5).