Fig. 4
POSTN increases monocytes migration and promotes M2 macrophages polarization. A THP-1 cell migration ability was determined by in vitro migration assay upon addition of the control SKOV3/pcDNA4 or SKOV3/POSTN conditioned medium with or without specific neutralizing antibody pre-treatment in the bottom well. The αvβ3 or αvβ5 integrin neutralizing antibody, as indicated, was added before collecting the conditioned medium. *p < 0.01, ***p < 0.001. B Flow cytometry analysis of THP-1 cells treated with the vector control or POSTN-overexpressing conditioned medium to measure CD68-positive, CD206-positive or CD80-positive populations (left, middle and right, respectively). **p < 0.01, ***p < 0.001. C Flow cytometry analysis of THP-1 cells treated with conditioned medium from the non-specific (N/S) shRNA control or shPOSTN SKOV-I6 cells to measure CD68-positive, CD206-positive or CD80-positive populations (left, middle and right, respectively). *p < 0.05, **p < 0.01. D, E qRT-PCR analysis of THP-1 cells co-cultured with SKOV3 cells transfected with a control or POSTN-overexpressing vector for 5 days to measure M2- or M1-associated cytokines expression change (D, E, respectively). The mRNA expression levels were compared based on the relative fold-change of each gene and the associated p values were calculated using a paired t test. *p < 0.05, **p < 0.01