library_construction_protocol: Medial prefrontal cortex (mPFC) tissues were harvested from four Npas2 KO mice and four wild-type littermate control mice at ZT20. To minimize individual biological variance and ensure sufficient nuclear yield, tissues from mice of the same genotype were pooled prior to dissociation.Nuclei were isolated using the Shbio Cell Nuclear Isolation Kit (Shibio, Cat# 52009-10) in conjunction with NP-40 lysis buffer to facilitate efficient membrane lysis while preserving nuclear integrity. The pooled tissues were homogenized according to the manufacturer's instructions. The resulting homogenate was filtered to remove cellular debris, and the nuclei were recovered through centrifugation. Single-nucleus libraries were then constructed using the 10x Genomics Chromium Next GEM Single Cell 3' Reagent Kit v3.1, followed by high-depth sequencing on the Illumina NovaSeq 6000 platform with a paired-end 150 bp (PE150) configuration.