CLOCK and BMAL1 drive rhythmic transcription of Atgl and Hsl via E-boxes. A: Maps of the 5′ regions of the genomic loci of murine Atgl (top) and Hsl (bottom) on chromosome 7 (Chr.7). Putative E-box enhancers are indicated by ovals. Black arrows depict the genomic sequences cloned for promoter studies.
Mutated E-boxes are indicated by gray arrowheads. B: Luciferase reporter assays in HEK293 cells for wild-type (WT) and mutated Atgl (left) and Hsl (right) promoters in response to cotransfection with Clock/Bmal1 and Cry1 (n = 3; ***P < 0.001 by one-way ANOVA with Bonferroni posttest). C: Time-dependent BMAL1 binding to Dbp, Atgl, and Hsl E-boxes and 500-bp downstream regions as identified by ChIP in WAT (n = 3/time point [*P < 0.05; **P < 0.01; ***P < 0.001 by two-way ANOVA with Bonferroni posttest]). All data are shown as means ± SEM.