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. 2014 Jun 17;5(14):5350–5368. doi: 10.18632/oncotarget.2110

Fig.6. Neutralizing Antibody Mediated Blockade of Eotaxin Prevented Oncostatin M Mediated Macrophage Chemotaxis and their M2-Polarization.

Fig.6

THP-1 derived macrophages derived macrophages were incubated with conditioned media from hypoxia primed (6hrs) or normoxic breast cancer cells (MDA-MB-231) CM for 24 hrs in presence of recombinant Oncostatin M either alone or in combination with anti-Eotaxin neutralizing antibody or isotype control antibody. Thereafter macrophage migration and phenotype switching was evaluated using Geimsa/DAPI staining and flowcytometry respectively. Representative giemsa-staining photomicrographs and Flow cytometry data depicting blockade of directional migration and M2-polarization of THP-1 derived macrophages. Quantification of macrophage migration chemotaxis was done by DAPI staining of migrated macrophages followed by counting of nuclei in five different fields of three replica wells. Numbers in parenthesis represent % M2-macrophage count obtained during flow cytometric analysis of three replica sets. Data presented as Mean±SEM; n=5; Symbols indicate statistical significance at p < 0.05 (*- #). Differences between values with matching symbol notation are statistically insignificant.