RNA-seq of p5 rat brain cd11b/c control
Cell pellets were processed by the Cancer Genomics Center at The University of Texas Health Science Center at Houston (CPRIT RP180734). Total RNA was isolated from cell pellets using the Qiagen RNeasy Mini Kit (#74104) and the RNase-free DNase Set (#79254) per the manufacturers instructions. Total RNA was quality-checked using the Agilent RNA 6000 Pico kit (#5067-1513) by Agilent Bioanalyzer 2100 (Agilent Technologies). RNA with an integrity number >7 was used for library preparation. Libraries were prepared with NEBNext Ploy(A) mRNA Magnetic Isolation Module (New England Biolabs; E7490L), NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, E7760L) and NEBNext Multiplex Oligos for Illumina (New England Biolabs, E6609S) per the manufacturers instructions. Quality of the final libraries was examined using the Agilent High Sensitive DNA Kit (#5067-4626) with the Agilent Bioanalyzer 2100, and library concentrations were determined by quantitative polymerase chain reaction (qPCR) using the Collibri Library Quantification kit (ThermoFisher, A38524500). Libraries were pooled evenly and analyzed using paired-end 75-cycle sequencing on an Illumina NextSeq 550 System (Illumina, Inc.) using the High Output Kit v2.5 (#20024907, Illumina, Inc.).