Flow cytometric analysis of tumors identifies a distinguishing TEM signature. (A) Flow analysis of some of the genes that were either differentially expressed or not differentially expressed between TEMs and TAMs. N202 mammary tumors grown subcutaneously in Tie2-GFP transgenic were made into single-cell suspensions and TEMs and TAMs gated as Tie2-GFP⁺Cd11b⁺7AAD⁻ and Tie2-GFP⁻Cd11b⁺7AAD⁻ cells, respectively. In agreement with the mRNA data, TEMs but not TAMs robustly and uniformly express Tlr4, Mrc1, Il4ra, and Cd163 (left panels). Genes not differentially expressed by quantitative PCR are similarly expressed by TEMs and TAMs (right panels). For each marker, at least 3 independent analyses were performed; for each analysis, at least 3 tumors were pooled together and made into single-cell suspensions. (B) Further phenotypic characterization of TEMs and TAMs. Note that TEMs do not express c-kit or granulocyte (Ly6G), inflammatory monocyte (Ly6C), DC (Cd11c), mast cell (FcϵRI), and megakaryocyte/platelet (Cd41) markers. For each marker, at least 3 independent analyses were performed.