Extended Data Fig. 2: Control chow and PLX chow feeding in neuronal activity during emergence from anesthesia.

a, Experimental timeline of control chow feeding and two-photon imaging of neuronal Ca²⁺ activity. Related to main Fig. 2. b–d, Neuronal Ca²⁺ signal amplitude (b), time active (c) and signal area (d) across experimental time points for control chow 3-weeks group and 6-weeks group(amplitude: Awake versus Isoflurane, p = 0.9676; Awake versus Emergence 15 min, p = 0.8172; Awake versus Emergence 30 min, p = 0.9713; Awake versus Emergence 45 min, p = 0.9911; Awake versus Emergence 60 min, p = 0.9530. time active: Awake versus Isoflurane, p > 0.9999; Awake versus Emergence 15 min, p = 0.9733; Awake versus Emergence 30 min, p = 0.9897; Awake versus Emergence 45 min, p = 0.9697; Awake versus Emergence 60 min, p = 0.9995. signal area: Awake versus Isoflurane, p > 0.9999; Awake versus Emergence 15 min, p > 0.9999; Awake versus Emergence 30 min, p = 0.9754; Awake versus Emergence 45 min, p = 0.9992; Awake versus Emergence 60 min, p > 0.9999). Solid lines represent the mean ± SEM from n = 4 mice. e–g, Neuronal Ca²⁺ signal amplitude (e), time active (f) and signal area (g) across experimental time points for before and after microglia ablation at each time point (amplitude: Awake versus Isoflurane, p = 0.9959; Awake versus Emergence 15 min, p < 0.0001; Awake versus Emergence 30 min, p < 0.0001; Awake versus Emergence 45 min, p < 0.0001; Awake versus Emergence 60 min, p = 0.0413. time active: Awake versus Isoflurane, p > 0.9999; Awake versus Emergence 15 min, p = 0.0069; Awake versus Emergence 30 min, p = 0.0009; Awake versus Emergence 45 min, p = 0.0601; Awake versus Emergence 60 min, p = 0.8613. signal area: Awake versus Isoflurane, p > 0.9999; Awake versus Emergence 15 min, p = 0.1259; Awake versus Emergence 30 min, p = 0.0013; Awake versus Emergence 45 min, p = 0.0239; Awake versus Emergence 60 min, p = 0.0657). Solid lines represent the mean ± SEM from n = 10 mice. h–j, Absolute value of neuronal Ca²⁺ signal amplitude (h), time active (i) and signal area (j) (amplitude: Awake versus Isoflurane, p = 0.9998; Awake versus Emergence 15 min, p = 0.7250; Awake versus Emergence 30 min, p = 0.7176; Awake versus Emergence 45 min, p = 0.9757; Awake versus Emergence 60 min, p > 0.9999. time active: Awake versus Isoflurane, p > 0.9999; Awake versus Emergence 15 min, p = 0.4355; Awake versus Emergence 30 min, p = 0.5141; Awake versus Emergence 45 min, p > 0.9999; Awake versus Emergence 60 min, p = 0.8270. signal area: Awake versus Isoflurane, p > 0.9999; Awake versus Emergence 15 min, p = 0.9626; Awake versus Emergence 30 min, p = 0.8873; Awake versus Emergence 45 min, p > 0.9999; Awake versus Emergence 60 min, p = 0.9442). Solid lines represent the mean ± SEM from n = 10 mice. Two-way ANOVA followed by Sidak post-hoc test (b–j); n.s., not significant; *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.

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