Supplementary Figure 3 from Cancer-Associated Adipose Tissue Promotes Breast Cancer Progression by Paracrine Oncostatin M and Jak/STAT3 Signaling

Upper left panel: Image representing a protein array of 507 proteins used for detection of soluble factors present in a mixture of CM^CAAT from two patients. Each protein is spotted in duplicate. The boxes show the positive controls. The brackets indicate adiponectin (1) confirming the adipose nature of CM^CAAT, GCSF (2), IL-6 (3), IL-8 (4), leptin (5), LIF (6), OSM (7). Upper right panels: western blot analysis of Y705 pSTAT3 of MCF-7 cells treated for 48h with control medium; neutralizing antibodies against hIL-6, hLIF and hIL-8 (1 µg/ml); rhIL-6, rhLIF and rhIL-8 (10 ng/ml); combination of the recombinant protein with the neutralizing antibody (same concentrations); western blot analysis of MCF-7 cells treated for 48h with CM^CAAT and neutralizing antibodies against hIL-6, hLIF and hIL-8 at indicated concentrations. Lower panel: Phase-contrast images of MCF-7 cells treated or 48h with control medium, rhIL-6, rhLIF and rhIL-8 at indicated concentrations, CM^CAAT and CM^CAAT combined with neutralizing antibodies against hIL-6, hLIF and hIL-8 at indicated concentrations (Scale bar is 100 µm).