Wt pico amplification kit

Macrophages (F4/80⁺ CD11b⁺ 2–5000 cells) were sorted from a pool of ipsilateral L4/L5 DRG of SNI WT and miR-21 cKO using a FACS Aria II sorter (BD Bioscience). Total RNA was prepared from the cell lysate. Each condition was represented by independently collected biological triplicates. Labeled cell extracts were processed for microarray analysis using the WT Pico Amplification kit (Thermofisher) and hybridized to Affymetrix Mouse 430V2 Arrays. MAS5 pre-processed data were generated in Expression Console (Thermofisher) and analyzed for differential gene expression using Transcript Analysis Console (Thermofisher) with a P value cutoff < 0.05 and two-fold change filter applied. Statistically significant differentially expressed gene list associated with each condition were further annotated and interrogated using MetaCore software (Reuters).

Macrophages (F4/80⁺CD11b⁺, 2–5,000 cells) were sorted from a pool of ipsilateral L4/L5 DRGs of SNI WT and miR-21–cKO mice using a FACSAria II sorter (BD Biosciences) or cultured PMs. Total RNA was prepared from the cell lysates. Each condition was represented by independently collected biological triplicates. Labeled cell extracts were processed for microarray analysis using the WT Pico Amplification kit (Thermo Fisher Scientific) and hybridized to Affymetrix Mouse 430V2 Arrays. The quality of cDNA and fragmented cDNA was assessed in an Agilent Bioanalyzer 2100. All analyses were performed in R (v4.2.0). Statistically significant differences between groups were determined using the affy R package (52 (link)). The parameters were set to RMA background correction and quantile normalization, with pm correct pmonly and amedianpolish. Significant differential expression was inferred based on a P value of less than 0.05. Enrichment for GO terms for individual comparisons was performed using the EnrichGO function from the clusterProfiler R package in Bioconductor. A P-value and q-value cutoff of 0.05 was used.