Fig. 2

LAG-2 regulation of L4/A lethargus sleep cannot be ascribed to cell fate changes. a Over-expression of LAG-2 in adult animals induces inappropriate sleep bouts. Animals were heat shocked and percentage displaying inappropriate sleep was determined after a 1 h recovery period at 20 °C. * denotes statistical significance at p < 0.001 versus hsp::empty based on student’s two tailed t-test; error bars represent the standard error of the mean. b Adults in inappropriate sleep bouts had increased arousal thresholds. Percent response of sleeping animals is reported for two independent trials, n = 20 animals in total for each genotype. * denotes statistical significance, determined as in panel A, of p = 0.02 versus hsp::empty; error bars represent the standard error of the mean. c Increasing lag-2 activity before and during L4/A lethargus restored normal sleep quantity. Animals raised at the restrictive temperature of 25.5 °C were shifted to 15 °C 4 h prior to L4 lethargus (illustration, left side of panel) lag-2(q420) is a temperature-sensitive, loss of function allele with decreased function at higher temperatures. Lack of statistical significance based on student’s two tailed t-test. Wild type n = 13 animals and lag-2(q420) n = 14 at 15 °C; wild type n = 15 animals and lag-2(q420) n = 19 at 25.5 °C. d Decreasing lag-2 activity before and during L4/A lethargus did not impact sleep quantity. Animals raised at the permissive temperature of 15 °C were shifted to the restrictive temperature of 25.5 °C for 4 h prior to L4 lethargus (illustration, left side of panel). Total time in sleep bouts is reported in minutes as in panel C. Wild type n = 13 lag-2(q420) n = 14 at 15 °C, wild type n = 10 and lag-2(q420) n = 14 at 25.5 °C. Statistical significance determined as in panel A