Figure 1. Protocol and timeline for circadian disruption, dietary changes, and stool collection.

A. Non-shifted mice were kept on a constant light:dark schedule for the entirety of the experiment whereas Shifted mice underwent once weekly light:dark inversion. Mice were maintained on a standard chow diet for the first 12 weeks of the study followed by a stool collection at the end of week 12. Mice were subsequently placed on a high-fat, high-sugar diet for 10 weeks and stool was collected at the end Week 22. Representative behavioral activity recording for a Non-shifted (B) and Shifted mouse (C). Activity is represented as dark areas in the actogram. These data demonstrate that non-shifted mice have stable behavioral patterns while the shifted mice have disrupted behavioral patterns. D. Expression of Per2 mRNA in the proximal colon one day after the week 22 stool collection demonstrates altered circadian expression in the intestine following the 22 weeks of phase shifting. Significant effect of light condition, (F_(1,58) = 5.84, p = 0.02), Zeitgeber Time (ZT) (F_(5,58) = 8.38, p<0.0001), and light condition x ZT interaction (F_(5,58) = 9.67, p = <0.0001). E. Activity onset phase distribution on the day of stool collection for standard chow-fed and (left) and high-fat, high-sugar diet (right). Individual mouse onset times relative to the light:dark cycle are depicted by the circles, and vector means depicted by the lines. There were no significant differences between the mean phase angle of entrainment between the shifted and non-shifted mice at the time of stool sample collection, but shifted mice exhibited a greater dispersal of phases in both diets (p<0.05).