Extended Data Fig. 3: RUVBL2 is essential for circadian rhythm of locomotor activity and can be stimulated by external cues.

a, Histograms of circadian periods of SCN-specific transgenic mice. n = 7, 8, 9, 11 biologically independent mice for AAV-WT, Δ151-152, Δ86-87/Δ350-352/Δ83 and GFP. b, Schematic illustration of generating the SCN-specific knockout mice. c, RUVBL2 expression in the SCN. d, Histogram of RUVBL2-positive cells in the SCN. n = 7 and 13 biologically independent mice for control and treat group, position where dense nuclei appeared was defined as 0 μm. e, Periodgram of SCN-specific Ruvbl2 knockout mice. n = 10 and 12 biologically independent mice for control and treat group. f, Left panel: histograms of circadian periods in transgenic mice. Right panel: actograms of mice and the corresponding periodogram. Gray line represents Ruvbl2^WT introduction in the SCN. n = 9 mice per group. g, The mRNA expression levels of Ruvbl2 in liver and muscle. Three technical replicates from a mixed sample of 5 mice. h, Statistical analysis of Per2 and Ruvbl2 expression in mouse liver, and blood glucose, G6P as a positive control in fasting-refed experiments. Three technical replicates from a mixed sample of three mice for gene expression experiment. n = 9 (ZT12), 6 (ZT15), 3 (ZT17) biologically independent mice for blood glucose. i, Expression of RUVBL2 and c-Fos in the SCN, light pulse stimulation at CT15, data were analyzed from 5 mice per group. All period data parameters were analyzed from ClockLab Analysis. Two-way ANOVA differential analysis with Dunnett’s multiple comparisons test (d). One-way ANOVA differential analysis with Šídák’s multiple comparisons test (f). Two-tailed multiple unpaired t-test differential analysis with Two-stage step-up (Benjamini, Krieger, and Yekutieli) method (h). Two-tailed t-test differential analysis (i). All graphs show mean ± s.e.m. Illustration in a created using BioRender (credit: Juan, C. https://BioRender.com/n10u764; 2024).

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