Fig 4.

A-C. Most Madeira cockroaches retained circadian rhythmicity (n = 8 of 12), but expressed weaker (n = 6 of 12) and/or desynchronized rhythms (n = 3 of 12) with more than one period(τ) after injections of Rm´per dsRNA. (A) Double-plotted running wheel activity of a Madeira cockroach in constant darkness 7 days before, and 29 days after the injection of Rm´per dsRNA (star). At day 36 of the locomotor activity recording the cockroach was sacrificed (cross) and qPCR was used to confirm the knockdown. Chi-square periodogram analysis of the number of running wheel turns indicated significant rhythmicity with a period of 24.08 h 1–7 days before Rm´per dsRNA injection (B) and 23.72 h 2–4 weeks after the injection (C). Already before injection the broad, two peaked distribution of significant rhythmicity indicated only loosely coupled oscillators controlling locomotor rhythms in this individual cockroach. Knockdown of Rm´per mRNA further dissociated underlying short and long rhythmic components (dashed lines), but did not completely delete rhythmicity over the course of 4 weeks.