title: GSM7795271: Sample_3_Neutrophils_APOE4_Spleen; Mus musculus; RNA-Seq
library_construction_protocol: Spleens or brain were mashed with the plunger of a syringe and treated with 1ml of ACK for 10min buffer to remove erythrocytes. Splenic cells were washed, pelleted and stained with the following rat antibodies: BV421 anti-CD45 (1:100, Biolegend, 103133), PE-Cy7 anti-mouse CD11b (1:300, eBioscience, 50-154-54), PerCP/Cy5.5 anti-mouse Ly-6C (1:300, Biolegend, 128012), Alexa Fluor 488 anti-CD115 (1:100, Biolegend, 135512) and Alexa Fluor 700 anti-Ly6g (1:100, Biolegend, 127621) in PBS with 2% FBS. After washing, CD115-CD11b+Ly6cintLy6g+ and CD115-CD11b+Ly6cintLy6g+/Tdt neutrophils were sorted using BD FACSAriaTM II (BD Bioscience). Smart-Seq2 libraries were prepared by the Broad Technology Labs and sequenced by the Broad Genomics Platform. cDNA libraries were generated from sorted cells using the Smart-seq2 protocol. RNA sequencing was performed using Illumina NextSeq500 using a High Output v2 kit to generate 2× 25 bp reads.