MCAO-V-4 (CRX1864492; CRA017573)
The microglial total RNA was extracted using the TRlzol reagent (15596026CN, Thermo Fisher). A total amount of 1 microg RNA per sample was prepared for the RNA sequencing. Sequencing libraries were generated using the Hieff NGS Ultima Dual-mode mRNA Library Prep Kit for Illumina (Yeasen Biotechnology). mRNA was purified using poly-T oligo-attached magnetic beads. The AMPure XP system purified the library fragments (Beckman Coulter). PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers, and Index (X) Primer. The libraries were sequenced on an Illumina NovaSeq platform to generate 150 bp paired-end reads.