FIG. 8.

Acute H₂O₂ and diamide treatment induce rapid PER2 degradation. (A) Western blot analysis of cell lysates of PER2::VENUS cells treated with H₂O₂ (60 μM–1 mM) or diamide (250 μM) for 15 min at the expected peak of PER2 expression. The graph depicts the PER2::VENUS levels relative to tubulin expression. (B) Experiment as in (A). Effects of acute oxidative stress (H₂O₂ or 250 μM diamide) for 15 min, at the normal PER2 peak, on PER2 stability, in the presence or absence of the inhibitor of protein synthesis CHX (10 μM). A similar experiment assessing PER2::LUC bioluminescence can be found in Supplementary Figure S9. CHX, cycloheximide.