ScRNA-seq and SnRNA-seq respectively for P3 and P22 mice brain

To explore the function of Rack7 in brain development, we deleted Rack7 in the CNS using a human glial fibrillary acid protein (GFAP) promoter-driven Cre (hGFAP-Cre), which is expressed at embryonic day (E) 13.5, and targeting multi-potential neural stem cell for neuronal and glial cells. Homozygous conditional deletion of Rack7 (hGFAP-cre:: Rack7fl/fl, referred as cKO) was verified by PCR and immunoblotting(compared with wildtype mice (hGFAP-cre:: Rack7+/+, referred to as WT). As RACK7 was identified as a transcriptional repressor, to obtain more precise insights into the consequences of RACK7 loss, we utilized high-throughput single-cell(sc) and single-nucleus(sn) RNA sequencing to examine the transcriptional landscapes of Rack7-cKO and wildtype brains. We extracted the cortical and hippocampal tissues from Rack7-cKO and wild-type mice at P3 (scRNA-seq) and P22(snRNA-seq), prior to processing with the 10X Genomics platform.