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Fig. 2 | Nutrition & Metabolism

Fig. 2

From: Ammonia-lowering activities and carbamoyl phosphate synthetase 1 (Cps1) induction mechanism of a natural flavonoid

Fig. 2

NOB modulates Cps1 mRNA and protein expression. a Total protein extracts were prepared from liver samples collected from the four diet/treatment groups of wild-type mice at the indicated circadian time points (n = 3). Western blotting analysis was performed using anti-CPS1 antibody. RC regular chow, HFD high-fat diet, Veh vehicle, NOB Nobiletin. The results shown are representative of three independent experiments. See Additional file 1: Figure S1A for quantitative analysis. b Immunohistochemical staining of CPS1 in liver sections from mice with the indicated diet and treatment at ZT2. c Real-time RT-PCR analysis of Cps1 in liver samples collected as in (a). Data are presented as mean ± SEM (n = 3). Two-way ANOVA with Bonferroni post-hoc tests shows significant statistical differences between HFD.Veh and other three groups (p < 0.0001). d Western blotting analysis of protein lysates of liver samples collected at ZT 6 and 18 from mice with the indicated diet and treatment (n = 3). HPD indicates high-protein diet. The images shown to the left are representative of three independent experiments. Quantitation of Western blots was carried out and the results, presented as mean ± SEM, are shown in the lower panel. Two-way ANOVA with Bonferroni post-hoc tests, RC vs. HPD, ***p < 0.001. e Real-time qPCR analysis was carried out using total RNAs extracted from the liver samples described in (d). The results are presented as mean ± SEM. Two-way ANOVA with Bonferroni post-hoc tests, RC vs. HPD, *p < 0.05

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