Figure 1.

Experimental design and validation. A) Overview of experimental design. A cohort of 10 male C57BL/6J mice was fasted for 24 hours, then split into fasted and re-fed groups (n=5 each). The fasted mice were immediately sacrificed, while the re-fed group was given access to food for 2 hours before sacrificing. Blood glucose measurements were taken from all mice prior to sacrifice. Livers were collected shortly after sacrifice, and used for both mRNA and ChIP-seq experiments. B) Blood glucose was significantly higher in re-fed animals. Error bars indicate SEM, *p < 1E-4, one-tailed Student’s t-test. C) Known fasting-induced gluconeogenic genes are significantly induced in the fasted state (white bars) relative to the re-fed state (black bars) by RT-PCR. Error bars indicate SEM, *p < 0.005, one-tailed Student’s t-test). D) Western blot analysis of mouse livers shows that CREB is strongly phosphorylated on Ser133 after a 24 hr fast, but not after 2 hour re-feeding. Western blot was re-probed with an antibody for total CREB to validate equal loading.