Figure 6.

Representative individual double-plotted actograms of ITP-KD, PDF-KD, and ITP/PDF-KD flies and controls in LD 12:12 followed by DD. ITP-KD flies (tim(UAS)G4>dcr2;itp-RNAi) (B) have significantly longer free-running periods in DD compared with the relevant controls (A). Many of the PDF-KD flies (tim(UAS)G4>dcr2;pdf-RNAi) were arrhythmic in DD (D). The still rhythmic individuals free ran with a short period (B) that was significantly different from the relevant controls (A). The majority of the ITP/PDF-double-KD flies (tim(UAS)G4>dcr2;itp-RNAi/pdf-RNAi) were arrhythmic (C, right actogram, D). The remaining flies showed several free-running components in DD (C, left actogram), the period of which was impossible to determine (F). Furthermore, all ITP/PDF-double-KD flies had a high activity level that was significantly different from all other genotypes (E). Black and white bars indicate the light regime in LD 12:12 (100 lux, 20°C). The control strains in D and E (dark gray bars) are in the following order from left to right: tim(UAS)G4>dcr2, dc2;itp-RNAi, and dcr2;pdf-RNAi. Error bars indicate SEM. Lowercase letters indicate statistical significances (different letters indicate p < 0.05). F, Characteristic periodograms for PDF-KD and ITP/PDF-KD flies. When only PDF was knocked down (top left periodogram), a significant period around 23 h was visible. In the ITP/PDF-double-KD flies, either no significant period was detectable (bottom left periodogram) or multiple (>3) periods appeared (right periodogram). The top right periodogram stems from the left actogram shown in C. A–C and F show individual actograms and periodograms, respectively. n ≥ 21 in D and E.