Figure 4
Inactivation of either Cn, NFAT1 or NFAT2 impairs the metastatic potential of 4T1 cells. (a) Weight of the lungs excised from killed mice, 2 weeks after intravenous injection of 5 × 105 4T1 cells. Note the significant decrease of the lung weight of mice injected with shCnB1-transduced cells (0.29±0.02 g, n=5), shNFAT1-transduced cells (0.16±0.02 g, n=5) or shNFAT2-transduced cells (0.21±0.01 g, n=3) cells versus control (LKO) 4T1 cells (0.40±0.02 g, n=5). Data are represented as mean±S.E.M. (b) Quantification of lung colonization after excision, fixation, paraffin embedding and H/E staining of lung sections of mice described in (a). Surface area of metastatic foci and surface of lungs were measured using ImageJ software. Data are represented as mean±S.E.M. (c) Representative pictures of lung sections as described in b after staining for cleaved caspase 3 and Hematoxilin/Eosin. Insets show a higher magnification of the underlined square. Note the brown CC3-positive cells specifically in shCnB, shNFAT1 and shNFAT2 metastatic foci. (d) Quantification of the number of apoptotic cells (CC3+) in metastatic foci of lungs, 15 days after intravenous injection of 4T1 cells. Data are plotted as fold increase compared with control and are the means of three independent tumors (n=3, data are represented as mean±S.E.M.)
