Fig. 3.

Effects of exogenously administration of IL-17A on cell migration, gene expression of pro-inflammatory cytokines and chemokines in MDA-MB-231 and MDA-MB-468 cell lines. MDA-MB-468/231 cells were cultured in a transwell system as described in method followed by administration of different doses of IL17A (0-, 1-, 10 ng/mL) in the lower chamber. After 4 h (for MDA-MB-231) or 8 h (for MDA-MB-468), the migratory cells were photographed, quantified (one-way ANOVA) (a). For Western blot analysis, cells (1 × 10⁶ / well) were cultured for 24 h in low serum medium, followed by another 24 h-culture, and then cells were harvested and probed with specific antibodies, quantified (one-way ANOVA) (b). mRNA transcripts of pro-inflammatory cytokines (TNF-α, IL-1β) and chemokines (CXCL2, CCL20) and COX2 were analyzed with real-time PCR (Mann–Whitney U test) (c). IL-17A gene expression including mRNA and protein level (d) was quantified by real-time PCR and ELISA assay (one-way ANOVA), respectively. Asterisk indicates a p value < 0.05