Figure 3. Transcriptional and phenotypic profile of M1 and M2a polarized macrophages.

(A) Venn diagram showing commonly and uniquely regulated probe sets found in M1 and M2a polarized macrophages. Probe sets that revealed a twofold or greater difference (|log₂ ratio| > 1.0) and rank product FDR <10% relative to M0 cells are shown. Data is averaged from 3–5 arrays for each subset. (B) Heatmap showing the fold change in expression of the 1891 differentially regulated probe sets identified in A. Genes are organized into unique and co-regulated gene clusters as indicated. (C) Annotated heat map showing the expression levels of select differentially regulated probe sets. For B and C the scale (–64.0 fold to +64 fold) represents the fold change relative to the mean normalized intensity value (log₂ ratio) in M0 cells (n=5). (D) q-PCR analysis of select gene expression in M0, M1, and M2 cells (n=3 for each gene). (E) Representative FACS analysis of M0, M1, and M2 macrophages (n=3) derived from bone marrow cells compared to PECs. (F) q-PCR analysis of Arginase 1 and NOS2 expression in PECs from mice pre-treated with IL-4 and IL-13 or PBS controls (n=4 for each group). (G) Representative flow cytometry analysis of F4/80 and YFP expression of recovered PECs, 48 hours after pre-treatment of Rag1^−/− C57BL/6 with YFP⁺ M2a macrophages and frequency of F4/80⁺YFP⁺ PECs (n=3). (H) q-PCR analysis of Arginase 1 and NOS2 expression in PECs from mice in (G). F4/80⁺ YFP⁺ cells are donor M2a macrophages while F4/80⁺ YFP⁻ cells are recipient mouse PECs.