A flow cytometry panel consisting of Lin-1, CD45, CD11b, F4/80, and CD274 (PDL1) was used for quantification and characterization of TAMs. Tumor cells were dissociated using a gentleMACS™ Tissue dissociator (Miltenyi) and tumor dissociation kit (Miltenyi) as recommended by the manufacturer. For intracellular staining of Vimentin and phospho-STAT3 (p-STAT3), cells were prepared using the Fix and Perm Kit as recommended by the manufacturer (Invitrogen). Antibody-capture beads (CompBeads, BD Biosciences) were used as single-color compensation controls. Cytometer calibration was performed daily by the use of rainbow fluorescent particles (BD Biosciences), after acquiring unstained and single-color control samples to calculate the compensation matrix. Data were analyzed using FCS Express software.

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