GSM5345208: KO_3; Mus musculus; RNA-Seq
Two days following CD40 agonist treatment, mouse livers were isolated and processed to generate single cell suspensions. Cells were stained for CD45.1 and CD45.2 and neutrophil-labeling markers, and sorted directly into Trizol and put on ice. RNA was extracted using TRIzol™ Plus RNA Purification Kit (Thermo Fisher) according to manufacturer's instructions. The Total-RNA samples were quantified using an Agilent Bioanalyzer 2100 instrument, with a corresponding Agilent Bioanalyzer RNA Pico assay. The resulting RIN (RNA Integrity Number) scores and concentrations were taken into account for qualifying samples to proceed. The samples were normalized to 1ng of input in 9uL (~111 pg/µL), and the mRNA was captured using oligo-dt primers as part of the Takara SMARTseq V4 Ultra Low workflow. The number of PCR cycles was increased to 13 to account for difficulties associated with neutrophil-derived RNA. A Bioanalyzer High Sensitivity DNA assay was run on the cDNA to evaluate success. cDNA synthesis, adapter ligation, and amplification were conducted subsequently with the Illumina NexteraXT Library Prep kit. Following amplification, residual primers were eluted away using Aline PCR CleanDx Beads. 
GEO Accession
GSM5345208