Protein samples were boiled in SDS sample buffer, separated by SDS-PAGE, and analyzed by immunoblotting with anti-CK2α (Santa Cruz Biotechnology, sc-6479), anti-CK2β (Calbiochem, 218712), anti-phospho CK2 substrate (Cell Signaling Technology, 8738), anti-phospho PKA substrate (Cell Signaling Technology, 9624), and anti–β-actin (Wako Pure Chemical, 017-24573) antibodies. For detection of BMAL1 S90-phosphorylation, an NIH-3T3 cell lysate was subjected to immunoprecipitation with anti-BMAL1 antibody followed by immunoblotting with anti-P-BMAL1-S90 and anti-BMAL1 antibodies (37). The protein concentration of each sample was measured by the Bradford method.

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