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Figure 4


Fig. 4. Essential role of NFAT3 in TNF-{alpha}-induced anchorage-independent growth of Cl41 cells. (A,C) Cl41-COX-2 mass1, Cl41-siNFAT3/COX-2 mass2 (1x104) were exposed to indicated concentrations of TNF-{alpha} in 1 ml of 0.33% MEM agar containing 5% FBS over 2.5 ml of 0.5% MEM agar containing 5% FBS in each well of a 6-well plate. After being cultured in a 37°C, 5% CO2 incubator for 3 weeks, the cell colonies were counted under microscopy, and colonies with more than 16 cells were scored. Each bar indicates the mean and s.d. from triplicate assays. (B,D) Schematic diagrams outlining the anchorage-independent cell growth in Fig. 4A,C. (E) The soft agar assay was performed as indicated in Fig. 3A, 2x104 cells were exposed to 20 U/ml of TNF-{alpha} in combination with various concentrations of CsA as indicated for 3 weeks. (F) Schematic diagram outlining the anchorage-independent cell growth in Fig. 4E.





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