GSM8447878: corticosterone group, scRNAseq; Mus musculus; RNA-Seq
The prefrontal cortical tissues of mice were freshly dissected and minced into small pieces on ice. The tissues were digested in RPMI 1640 (Thermo Fisher Scientific) / 0.1mg/ml Liberase TL (Roche) / 20g/ml DNase I (Sigma-Aldrich) / 10mM HEPES / 3% heat-inactivated fetal bovine serum (HI-FBS; Sigma-Aldrich) at 37oC for 15 min. The tissues were mashed through a 70-m cell strainer and centrifuged at 500 g for 5 min. The cells were re-suspended in PBS / 2% HI-FBS / 10mM EDTA-Na (pH 8.0) and stained with PE/Cyanine7 anti-mouse CD45 (BioLegend, #103114, RRID:AB_312979) and FITC anti-mouse/human CD11b (BioLegend, #101206, RRID:AB_312788). FACS-stained cells were processed on the Beckman MoFlo Astrios EQ, and microglia were sorted as CD45low CD11b+ by the common standard. The scRNA-seq libraries were constructed according to the protocol of the GEXSCOPE® Single Cell RNA Library Kits (Singleron) . Individual libraries were diluted to 4 nM, pooled, and sequenced on Illumina novaseq 6000 with 150 bp paired end reads.