Fig. 4

Decreased Notch signaling results in DAF-16 dependent increased sleep. a lin-12 Notch receptor loss results in daf-16-dependent increased sleep. Animals reared at 25 °C and assayed at 22 °C. lin-12(n941) is a complete loss of function allele; daf-16(mu86) is a partial loss of function allele. Box shows two middle quartiles, horizontal line indicates mean, bars represent maximum/minimum; * denotes statistical significance by two-tailed student’s t-test with p ≤ 0.001 versus wild type. Wild type n = 8, daf-16(mu86) n = 10, lin-12(n941) n = 10, daf-16(mu86);lin-12(n941) n = 12. b Decreased glp-1 Notch receptor function results in daf-16-dependent increased sleep. Parents shifted to 25 °C; F1 progeny assayed at 22 °C. glp-1(e2141) causes decreased function at higher temperatures; Statistical analysis as in panel A. *p ≤ 0.01 versus control. Wild type n = 14, daf-16(mu86) n = 13, glp-1(e2141) n = 14, daf-16(mu86);glp-1(e2141) n = 17. c Decreased daf-16 function did not ameliorate arousal threshold defects in glp-1 animals. Percent response reported for two independent trials; n = 20 animals for each data point with error bars for SEM. p = 0.11 for wild type versus glp-1(e2141) by t-test, but note that one way ANOVA suggests no statistical significance these genotypes or between glp-1(e2141) and daf-16(mu86);glp1(e2141). d Increased sleep quantity in lin-12 gain of function animals is daf-16-independent. Animals reared at 15 °C, and L4 stage animals assayed at 22 °C for total time in sleep bouts. lin-12(n137n460) is a temperature-sensitive, gain of function allele with increased function at lower temperatures. Statistical analysis as in panel A;*p ≤ 0.001 versus wild type. Wild type n = 8, daf-16(mu86) n = 10, lin-12(n137n460) n = 10, daf-16(mu86);lin-12(n137n460) n = 12. e Increased arousal thresholds in lin-12 gain of function animals are daf-16-independent. Statistical analysis as in panel C. Four independent trials, with n = 40 for each data point. p ≤ 0.001 versus wild type. No statistical significance found by student’s two-tailed t-test for lin-12(n137n460) versus double mutant