SLOB protein cycles in both LD and DD. (A) Canton S flies were entrained in LD for three days and collected at six time points over a 24 hour period. Fly head lysates were prepared and an anti-SLOB antibody was used to identify the 58 kDa band representing SLOB on the top western blot. The bottom blot was probed for MAPK, the protein level of which does not cycle  and was used as a loading control. SLOB oscillation has a trough at ZT 2 and a peak between ZT 10–14. The graph below represents the mean ± SEM for each time point from a minimum of three independent experiments. (B) Canton S flies were entrained in LD for three days and transferred to constant darkness. They were collected on the second day of DD at six time points, and SLOB cycling was determined as in (A).