FIGURE 5.

Differential expression of selected core circadian clock genes and NR1D1 protein in ileum. RT-qPCR analysis of circadian clock genes in male (A) and female (B) rats from study A. (C) Representative Western Blot images of NR1D1 in female ileum (left), and densitometric analysis of three replicates of Western blot analyses and of eight biological replicates from each study group (right). When normalized for HPRT protein expression, there was no significant difference between the study groups (D) Immunostaining of NR1D1 protein (green) in small intestinal paraffin embedded tissue sections demonstrates nuclear localization in intestinal epithelial cells (representative image). Wheat germ agglutinin (WGA) immunostaining (red) was used for outlining cell borders and mucus; the blue color indicates staining of nuclei by 4’,6-diamidino-2-phenylindole (DAPI). ^∗P < 0.05; ^∗∗P < 0.01; ^∗∗∗P < 0.001; Mann–Whitney U-test.