A macrophage-tumor cell cross-talk within Amppos tumors.

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A macrophage-tumor cell cross-talk within Amppos tumors. A macrophage-tumor cell cross-talk within Amppos tumors. A, qPCR analysis of Hgf mRNA in WT livers, Ampneg, and Amppos tumors. The cross line signifies geometric mean (**, P < 0.001). B, representative photomicrographs of IHC for HGF on Ampneg and Amppos tumors. Scale bar, 100 μm. C, representative confocal microscopy images of Amppos tumor immunostained for vWF, F4/80, and HGF. Hoechst 33342 marks nuclei. Scale bar, 40 μm. D, representative cytospin preparations of macrophage and hepatocyte fractions from Mdr2−/− livers. E, qPCR analysis of hepatocyte and macrophage fractions (n = 11 different mice), isolated from livers of Mdr2−/− mice. Bars represent geometric mean (**, P < 0.001; ***, P < 0.0001). F, representative photomicrograph of IHC for the VEGFR KDR in Ampneg and Amppos tumors. Note that the expression of KDR is confined to endothelial and stromal cells. Scale bar, 100 μm. G, mRNA qPCR analysisfor the VEGFRs Kdr and Flt1 and the macrophage and endothelium markers Msr1 and Cd105 on tissue lysates from the indicated groups. Each dot represents a different tumor. The cross line represents geometric mean (***, P < 0.0001). H, peritoneal macrophages were cultured under serum-free conditions followed by 8 hours of exposure to recombinant murine VEGF-A (100 ng/mL). Hgf expression was measured by qPCR. Bars represent geometric mean; n ≥ 3; *, P < 0.05. Elad Horwitz et al. Cancer Discovery 2014;4:730-743 ©2014 by American Association for Cancer Research