Figure 3. Spatially oriented and graded responses of per-short phospho-cluster site mutants on clock speed.

(A to E, left panels) Flies bearing different dper transgenes (indicated at top of panels) in a wper⁰ genetic background were collected at the indicated times (top of panels) and head extracts analyzed for dPER by immunoblotting in the presence of αHA. (A to E, right panels) The corresponding locomotor activity profiles of the transgenic flies in LD are shown. The second and third days worth of activity data during LD were averaged, generating the 24-hr profiles shown in the panels. The free-running periods (τ) are also shown. Arrow represents the timing of the clock-controlled evening peak. Vertical bars represent the activity recorded in 15-minute bins during times when the lights were on (grey bars) or off (black bars). ZT0 is lights-on, whereas ZT12 is lights-off. Horizontal bars at bottom; open, lights-on; closed, lights-off. (F) dPER signal intensity (Y-axis) was normalized by setting the peak intensity value for each dPER variant to a value of 1. Two independent experiments were used to generate the average and error bars represent SEM. See Table S1 for supporting data.