Fig. 3
Alterations in lag-2 Notch signaling disrupted sleep bouts during C. elegans L4/A lethargus. a Increased sleep in animals with decreased lag-2 function was dependent on daf-16 function. Parental animals were shifted from 15 °C to 25.5 °C. F1 progeny were selected at L4 stage and assayed at 25.5 °C. Box shows two middle quartiles, horizontal line indicates mean, bars represent maximum/minimum; * denotes statistical significance with p < 0.05 by student’s two tailed t-test. Wild type n = 26 animals, daf-16(mu86) n = 14, lag-2(q420) n = 33, daf-16(mu86);lag-2(q420) n = 14, daf-16(mgDf50null) n = 10, daf-16(mgDf50null);lag-2(q420) n = 15. b Arousal threshold defects caused by decreased lag-2 function were not dependent on daf-16 function. Percent response is indicated from at least four independent trials, n = 40 animals for each data point. Statistical significance was assessed by one way ANOVA followed by a Tukey post hoc-test; error bars respresent the SEM. *p < 0.05 for comparison of wild type versus lag-2(q420) and wild type versus daf-16(mu86;lag-2(q420). No statistical significance between lag-2(420) and daf-16(mu86;lag-2(q420). c Complete loss of daf-16 function did not alter sleep bout arousal thresholds in animals with decreased lag-2 function. Statistical analysis and presentation indicated as in Panel B. * p ≤ 0.05 for comparison of wild type and lag-2(q420). No statistical significance between lag-2(420) and daf-16(mgDf50;lag-2(q420). d In animals with decreased lag-2 function, daf-16 function was required in neurons for increased sleep. DAF-16 was expressed under the control of a neuron-specific or muscle-specific promoter in daf-16(mgDf50) lag-2(q420) animals. Statistical analysis and presentation represented as in Panel A. * indicates statistical significance with p ≤ 0.01 for comparisons to control. Animals tested: daf-16(mgDf50) n = 10, lag-2(q420) n = 33, daf-16(mgDf50);lag-2(q420) n = 15, Punc-119: daf-16(mgDf50) n = 14, Punc-119:daf-16(mDf50);lag-2(q420) n = 15, Pmyo-3:daf-16(mgDf50) n = 13, Pmyo-3:daf-16(mgDf50);lag-2(q420) n = 10