Fig. 2

Monocytes maintain M2-like polarization and experience significant changes in the gene expression after APS processing. a Mean fluorescence intensity (MFI) of hallmark macrophage polarization markers CD80 (M1) and CD163 (M2) and MHC-II in classical monocytes (CD45⁺CD⁻CD15⁻CD14⁺CD16^dim) and non-classical monocytes (CD45⁺CD3⁻CD15⁻CD14^dimCD16⁺). b Flow cytometry results of CD163⁺ cell concentration in classical monocytes and CD163⁺ and double negative (Neg⁻) in non-classical monocyte in APS and WBCs. Individual donors are denoted as circles for APS and triangles for WBCs. c Flow cytometry plots of classical and non-classical monocytes subtyping based on CD80 and CD163 in APS and WBCs. Phenotype plots of the percentage of CD80⁺, CD163⁺, CD80⁺CD163⁺, and double negative (Neg⁻) in classical and non-classical monocytes. d Flow cytometry plots of MHC-II⁺ classical monocytes and non-classical monocytes and quantification of the percentage of MHC-II⁺ in classical and non-classical monocytes in APS and WBCs, data are mean with ± SD. e Differential expression of the top 50 significantly upregulated genes in APS-sorted monocytes compared to WBCs-sorted monocytes (CD45⁺CD3⁻CD14⁺CD15⁻CD11c⁺CD11b⁺) from Nanostring gene expression assay. f Gene Ontology analysis of Nanostring gene expression data using MySigDB REACTOME Gene sets, p_adj < 0.05. g Gene set enrichment analysis (GSEA) using Nanostring Annotations v46. For all bar graphs, data are mean with ± SD. Multiple unpaired t-test without correction for multiple comparisons with set p-value threshold, alpha = 0.05 for b. *p < 0.05