Fig. 6

NOB restored Cps1 and Cebp mRNA and protein levels in a clock-dependent manner. a Liver samples were collected at the indicated Circadian Times (CT) from regular chow-fed WT and Clock ^Δ19/Δ19 mutant (Clk) mice (n = 3) in constant darkness (DD) when the circadian clock free runs devoid of light cue. CT0 corresponds to the onset of subjective day. Western blotting was performed using total protein extracts from the liver samples with the indicated antibodies. See Additional file 1: Figure S3 for quantitative analysis. b Real-time RT-PCR analysis of Cps1, Cebpa and Cebpb was carried out using total RNAs extracted from the liver samples described in (a). Data are presented as mean ± SEM. Two-way ANOVA with Bonferroni post-hoc tests, WT vs. Clk, Cps1, Cebpb p < 0.01; Cebpa p < 0.05. Student’s t-test, WT vs. Clk, Cps1: CT4, *p < 0.05; Cebpa: CT8, *p < 0.05; Cebpb: CT8, **p < 0.01. c Western blot analysis of CPS1 protein levels in liver samples collected at ZT8 and ZT20 from WT and Clk mice subjected to the indicated diet and treatment. RC regular chow, HFD high-fat diet, Veh vehicle, NOB Nobiletin. Two-way ANOVA with Bonferroni post-hoc tests, WT vs. individual Clk groups, *p < 0.05