Figure 1

Nuclear factors of activated T cells 5 (NFAT5) dictates the invasiveness of breast cancer cells. (A) qRT-PCR (upper panel) and western blotting (lower panel) assays for NFAT5 expression in breast cancer cell lines. Relative levels compared with the normal human mammary epithelial MCF-10A cells were plotted for qRT-PCR. (B) qRT-PCR and western blotting assays of MDA-MB-231 cells after transfection with control (100 nM) or NFAT5-targeted siRNAs. siNFAT5-2 was used thereafter due to its high efficacy in posttranscriptionally suppressing NFAT5 expression. (C, D) Transwell (C) and wound-healing (D) assays of cells transfected with scrambled or NFAT5-targeting siRNA. (E) MDA-MB-231 cells were modified with lentiviruses expressing scrambled or NFAT5-targeting shRNAs, and were injected into the tail vein of nude mice (n =10). The formation of lung metastases were monitored and representative H&E-stained lung tissues are shown. (F) qRT-PCR assay of 30 breast cancer samples (13 with metastases) and normal mammary tissues from four women undergoing breast reduction surgery. NFAT5 levels normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are plotted. All data are representative photographs or represent the mean ± SD of three replicates. *P <0.05, **P <0.01. VEGF-C: vascular endothelial growth factor C.