Figure 5.

Quantification of CO, HO-1, and the DNA/protein complexes in the liver of the hemoCD1-treated mice. The mice were housed for two weeks under a 12 h light/dark (LD) cycle (lights on at 7:00 and light off at 19:00) until the day before the experiments. The mice were then housed under constant dark (DD) conditions during the observations. (a) Endogenous CO levels were measured by the assay using hemoCD1³⁵. (b) HO-1 levels were ascertained by western blotting with an anti-HO-1 antibody at clock time 15.0 (first phase), 17.5 (second phase), and 19.5 (third phase). (c,d) ChIP assays were conducted using anti-NPAS2 and anti-CLOCK antibodies to measure the corresponding DNA complexes at clock time 14.8 (first phase), 17.5 (second phase), and 19.5 (third phase). PBS and Fb-hemoCD1 were used as controls. Each bar represents the mean ± SE (n = 3 mice per group). Asterisk denotes statistical significance (^*P < 0.05, ^**P < 0.01), as compared to the controls.