FIG. 5.
PER is coimmunoprecipitated with DBT from fly head extracts of wild-type, dbtS, and dbtL genotypes. Flies were collected at ZT21 (9 h after lights out in flies entrained to a 12-h LD cycle), and extracts were prepared. These were either electrophoresed directly (“total extract”) or subjected to immunoprecipitation with an antibody to the C terminus of DBT. After immunoprecipitation, the gammabind G-Sepharose with bound antibody-antigen complexes was resuspended in SDS-PAGE denaturation buffer, and an amount of the immunoprecipitate corresponding to three times the amount of extract analyzed in the first two lanes was subjected to immunoblot analysis for the detection of PER. Visualization of PER was accomplished with chemiluminescence. The genotype labels are described in the legend for Fig. 2. e, a mock immunoprecipitation with fly head extract only (no antibody); a, a mock immunoprecipitation with antibody only (no fly head extract).