FIG. 6.

mPER3 cannot participate in rhythmic assembly of clock protein complexes. (A) mPER complexes in wild-type (wt) and mPer double-mutant mice. Liver extracts from wild-type and mPer double-mutant mice collected at CT06 and CT18 were immunoprecipitated with antibodies to mPER1, mPER2, and mPER3 (P1, P2, or P3). The immune complexes were Western blotted for mPERs, mCRYs, and CKIɛ. (B) Interaction between CLOCK-BMAL1 and mCRY is abolished in mPer1/2 double-mutant mice. Liver extracts from wild-type (wt) and mPer double-mutant mice collected at CT06 and CT18 were subjected to IP with anti-CLOCK antibody. The immune complexes were Western blotted for CLOCK, mCRY1, and mCRY2 (left panels). Straight liver extracts were Western blotted for mCRY1 and mCRY2 (right panels) to show that mCRYs are readily detectable in mice of each genotype.