TIM-CLKGR flies display low amplitude circadian protein oscillations.

<p><b>A.</b> VRI levels oscillate in TIM-CLKGR flies with lower amplitude. A representative VRI Western blot from TIM-CLKGR and control (<i>tim-gal4/+</i> and <i>UAS-ClkGR/+</i>) fly heads. We collected the samples at 6 time points during the first day in constant darkness (DD1). <b>B.</b> Quantification of VRI oscillations in TIM-CLKGR flies. Western blot staining intensity (Quantification using ImageJ), normalized to tubulin. Average values of three independent biological repeats. Error bars represent SEM. One way ANOVA was performed to determine the statistical significance of the different between VRI levels of TIM-CLKGR and control flies, at CT 7 and CT11 *<i>p</i><0.05. <b>C.</b> Expression of CLKGR decreases the amplitude of VRI oscillation in circadian neurons. VRI (green) and PDF (red) immunostaining of the small Lateral Neurons ventral (sLNvs) from TIM-CLKGR and control (<i>tim-gal4/+</i>) fly brains. We collected samples at four time points during the first day in constant darkness. Pictures from one representative experiments are shown. <b>D.</b> Quantification of VRI Immunostaining in the sLNvs in constant darkness. Values are average staining intensity of at least 5 brains from each time point. Error bars represent SEM. Wilcoxon test was performed to determine statistical significance. **<i>p</i><0.01.