TIM-CLKGR flies display impaired function of peripheral oscillators.

<p><b>A.</b> Eclosion circadian gating is absent in TIM-CLKGR flies and in flies that express CLKGR under P{GawB}Mai60 driver (MAI60-CLKGR). The Mai60 driver express predominantly in the prothoracic gland. The eclosion ratio is calculated by determining the portion of flies that emerged in two hours intervals over the total amount of flies that emerged in 24 hours. The experiments were performed in TIM-CLKGR and MAI60-CLKGR flies and for the control lines UAS-CLKGR (UAS-<i>ClkGR/+</i>), TIMGAL4 (<i>tim-gal4/+</i>) and MAI60GAL4 (flies that carry the P{GawB}Mai60 insertion). Values are the means of 3 or 4 biological repeats (20 to 32 flies in each repeat) Error bars represent SEM. One-way Anova was performed to determine statistical significance of the differences between timepoints, for TIMGAL4, UAS-CLKGR and MAI60GAL4. <i>p</i><0.01. Experiment was performed in LD conditions <b>B.</b> Total sleep is not affected in TIM-CLKGR flies. Male flies were kept in 12∶12 LD conditions. Sleep was measured for 5 days. Sleep data was analyzed using pySolo software. Fly strains: TIM-CLKGR (n = 35), TIM-CLKGR(X2) (n = 32) and control flies TIMGAL4 (<i>tim-gal4/+</i>,n = 28) and UAS-CLKGR (UAS-<i>ClkGR/+</i>,n = 31). Values represent means and errors bars represent SEM. <b>C.</b> Expression of CLKGR leads to a dose dependent increase in the number of sleep episodes during the dark phase. Experimental conditions and genotypes as described in B. One-way Anova was performed to determine statistical significance of the differences between fly strains. <i>p</i><0.0001. <b>D.</b> Expression of CLKGR leads to a dose dependent decrease in the average length of the sleep episodes during the dark phase. Conditions and genotypes are as described in B. One-way Anova was performed to determine statistical significance of the differences between fly strains, <i>p</i><0.0001.