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 CMCAAT 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 CMCAAT, 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 CMCAAT 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, CMCAAT and CMCAAT combined with neutralizing antibodies against hIL-6, hLIF and hIL-8 at indicated concentrations (Scale bar is 100 µm).