Extended Data Fig. 2: Effect of Gi-DREADD activation on sleep in female and male mice, early light phase, dark phase, EEG power spectra within each state, and control experiments in mice without Gi-DREADD.

a, b, Summary of chemogenetic experiment, analyzed separately for female and male mice. Shown are the percentages of time in each brain state following CNO and saline injection (mean ± s.e.m.; a, n = 3 mice; two-way ANOVA with Bonferroni correction; NREM: P_treatment = 0.015; P_time < 0.0001; ^*P_1h = 0.017; ^**P_1.5h = 0.0089; Wake: P_treatment = 0.016; P_time < 0.0001; ^*P_1.5h = 0.015; REM: P_treatment = 0.36; P_time < 0.0001; b, n = 5 mice; two-way ANOVA with Bonferroni correction; NREM: P_treatment = 0.0001; P_time < 0.0001; ^**P_1h = 0.0057; Wake: P_treatment = 0.0002; P_time < 0.0001; ^**P_1h = 0.0092; REM: P_treatment = 0.21; P_time < 0.0001). c, Summary of chemogenetic experiment performed between Zeitgeber time (ZT) 1 (8:00 am) and ZT6 (1:00 pm) (mean ± s.e.m.; n = 8 mice: 4 female and 4 male; two-way ANOVA with Bonferroni correction; NREM: P_treatment = 0.0004; P_time < 0.0001; ^*P_1h = 0.032; ^**P_2.5h = 0.0014; Wake: P_treatment = 0.0007; P_time < 0.0001; ^***P_2.5h = 0.0002; REM: P_treatment = 0.40; P_time < 0.0001; ^*P_2.5h = 0.047). d, Percentage of time in each brain state within 3 h after CNO or saline injection. Each circle indicates data from one mouse (mean ± s.e.m.; n = 8 mice). ^***P < 0.001 (paired two-tailed t-test; ^***P_NREM = 0.0005; ^***P_Wake = 0.0006). e, f, Mean episode duration (e) and episode number per hour (f) for each brain state within 3 h after CNO or saline injection. Each circle indicates data from one mouse (mean ± s.e.m.; n = 8 mice). ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 (paired two-tailed t-test; e, ^***P_NREM = 0.0003; f, ^**P_NREM = 0.0099; ^*P_Wake = 0.010). g, Summary of chemogenetic experiment in dark phase (beginning at ~8:00 pm). Shown are the percentages of time in each brain state following CNO and saline injection (mean ± s.e.m.; n = 8 mice; two-way ANOVA with Bonferroni correction; NREM: P_treatment = 0.020; P_time = 0.012; ^*P_0.5h = 0.016; Wake: P_treatment = 0.040; P_time = 0.0039; ^*P_0.5h = 0.014; REM: P_treatment = 0.89; P_time < 0.0001). h, Percentage of time in each brain state within 3 h after CNO or saline injection. Each circle indicates data from one mouse (mean ± s.e.m.; n = 8 mice). ^**P < 0.01 (two-tailed Wilcoxon signed rank test; ^**P_NREM = 0.0078; ^**P_Wake = 0.0078). i, j, Mean episode duration (i) and episode number per hour (j) for each brain state within 3 h after CNO or saline injection. Each circle indicates data from one mouse (mean ± s.e.m.; n = 8 mice). ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001 (i, paired two-tailed t-test, ^****P < 0.0001; j, paired two-tailed t-test; ^***P_NREM = 0.0010; ^***P_Wake = 0.0012). k, l, Comparison of normalized EEG power spectra within each brain state between saline and CNO sessions during the light phase (k) or dark phase (l), averaged across 8 mice. Shading, ± s.e.m. The EEG spectra of each session was normalized by the total power between 0 and 25 Hz before averaging. m, Percentages of time in each brain state following CNO or saline injection in Tmem119-CreERT2 control mice without Gi-DREADD expression (mean ± s.e.m.; n = 6 mice). n, Changes in each brain state induced by chemogenetic manipulation (difference between CNO and saline injections, averaged across 3-h after injection) in mice without Gi-DREADD (control) or with Gi-DREADD (treatment during light phase (ZT1 – ZT6 and ZT6 – ZT11) or dark phase). Each circle indicates data from one mouse (Control, n = 6 mice; ZT1-ZT6, n = 8 mice; ZT6-ZT11, n = 8 mice; dark phase, n = 8 mice); error bar: ± s.e.m. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 (One-way ANOVA with Holm-Šídák’s test; NREM, P = 0.0005; ^**P_ZT1-ZT6 = 0.0034; ^***P_ZT6-ZT11 = 0.0007; ^**P_dark = 0.0013; Wake, P = 0.002; ^*P_ZT1-ZT6 = 0.019; ^**P_ZT6-ZT11 = 0.0020; ^**P_dark = 0.0064; REM, P = 0.49).

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