Importance of Dynamic Interactions for Circadian Rhythmicity.

<p>(A) Systematic RNAi-mediated silencing of circadian clock core and regulatory components. RNAi constructs were lentivirally delivered into U2OS cells harboring a <i>Bmal1</i>-promoter luciferase reporter and oscillation dynamics were monitored for several days (see also <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#pgen-1003398-g003" target="_blank">Figure 3</a>). Circles represent the difference in period (± s.e.m.; n = 3 independent experiments) relative to non-silencing controls (n>10) for two RNAi constructs (if available). Filled circles show additional amplitude and/or damping phenotypes. Cells were classified as arrhythmic (ar) if the fit to a cosine function resulted in a low correlation coefficient (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#pgen.1003398.s013" target="_blank">Text S1</a>). Period deviations of more than 2 hours are given (see also <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#pgen.1003398.s004" target="_blank">Figure S4A</a>). (B) Systematic overexpression of circadian clock core and regulatory components. Experiments were performed with lentivirally delivered overexpression constructs as described in (A) (see also <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#pgen.1003398.s013" target="_blank">Text S1</a>). <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#s2" target="_blank">Results</a> of three independent experiments (± s.e.m.) are given (also see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003398#pgen.1003398.s004" target="_blank">Figure S4B</a>). (C) Correlation of circadian phenotype with number of dynamic interactions. The combined phenotypic score from silencing and overexpression experiments is significantly different for components with many dynamic interactions (≥5) compared to those with few (<5) (t-test: ** p<0.01; Mann Whitney test: ** p<0.01).