Additional file 2: of Ibuprofen supports macrophage differentiation, T cell recruitment, and tumor suppression in a model of postpartum breast cancer

Figure S2. RNA Seq Analysis of Macrophages and Monocytes. a Involution D6 mammary glands were digested into single cell suspension and stained for characterization of F4/80+ populations by flow cytometry. Gates based upon F4/80 expression (orange F4/480-, blue-F4/80+ and red F4/80++) were populated into histogram overlays for comparison of lineage marker expression (CD45, CD11b, Ly6C) and activation (CD70, CD86, CD80) between F4/80+ and F4/80++ groups. b Principal component analysis of RNAseq data from involution day 6 mammary gland associated F4/80 low monocytes vs F/480 high macrophages with and without ibuprofen (IBU). c Complete gene lists for differentially expressed genes in macrophages (Mac) and monocytes (Mono) with and without ibuprofen treatment. Genes more highly expressed without ibuprofen are in red, while those more highly expressed with ibuprofen are in blue. d Examples of GSEA for transcription factor related gene pathways. Analysis by GSEA in which gene sets are composed of genes enriched in response to experimental overexpression of transcription factors (LEF-1) are annotated to have canonical transcription factor binding sites proximal to the indicated gene (STAT5). (TIF 9442 kb)