Transcriptomic analysis of WT and POLG mutator macrophages

The goal of this study was to characterize gene expression profiles of wild-type (WT) and POLG D257A/D257A mutator (POLG mutator) macrophages at rest and 6 hours after lipopolysaccharide (LPS) challenge. Both bone marrow-derived (BMDM) and thioglycollate elicited, peritoneal macrophages (PerMac) were analyzed. After 7 days of differentiation for BMDMs or 4 days post interperitoneal thioglycollate injection, macrophages were plated and stimulated with 200ng/ml LPS for six hours. Total cellular RNA from WT and POLG BMDMs and PerMacs was prepared using the Quick-RNA microprep kit (Zymo Research) and subjected to library preparation and RNA sequencing at the Experimental Genomics Core of the Texas A&M Institute for Genome Sciences and Society. FASTQ files were normalized, processed, and analyzed using BaseSpace Sequence Hub (Illumina). In brief, the STAR algorithm of RNAseq Alignment V2.0.0 software was utilized to align the results to the Mus musculus/mm10 (RefSeq) reference genome, then RNA-seq Differential Expression V1.0.0 software was used to obtain differential gene expression values and determine statistically significant changes in POLG mutator macrophages relative to WT.