Fig. 5

POSTN-overexpressing SKOV3 cells induces chemotactic cytokines expression in vitro and increases TAM in xenograft tumors. A Immunohistochemistry staining of POSTN and murine F4/80 (a marker of macrophages) molecules with respective antibodies using orthotopic xenografts tumor sections derived from SKOV3 cells transfected with control or POSTN-expressing vector. Scale bars = 100 μm. B Quantification of F4/80 staining intensity by ImageQuant under microscopy. **p < 0.01. C Conditioned media were collected from SKOV3 cells stably transfected with pcDNA4 (Ctrl) or pcDNA4/POSTN (POSTN) for cytokine array analysis. The cytokine expression was quantified by Image J software. D Cytokine mRNA expression was measured by qRT-PCR in SKOV-I6 cells transfected with either of the lentivirus-delivered shRNAs (shRNA#1 and shRNA#2) plasmid. Non-specific shRNA was used as the control (SC). E The effect on cytokine mRNAs production was detected by qRT-PCR after treatment with TPCA-1, an inhibitor of the NF-κB pathway (1 μM) or SN-50 (18 μM), an inhibitor against NF-κB nuclear translocation, for 48 h in control (white bars) or POSTN-overexpressing SKOV3 cells (black bars). DMSO (0.1 μL/mL) or water (solvent for SN50, labelled as UN) was used as the control. The bar graph represents the mean fold change ± SD from three independent experiments