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. 2016 Nov 28;36(20):2835–2845. doi: 10.1038/onc.2016.434

Figure 6.

Figure 6

NFAT3 phosphorylation is responsible for cell transformation, proliferation and tumor growth. (a) HaCaT cell lysates were examined using anti-NFAT3 or anti-phosphorylated NFAT3 (Ser259). (b) Stable HaCaT cells expressing wildtype or mutant NFAT3 were transiently transfected with the 3xNFAT3-luc reporter to access NFAT3 luciferase activity. Results are shown as relative luciferase units normalized to the luciferase activity of the mock control. The results were obtained from triplicate experiments and are shown as mean values±s.d. (c) The MTS assay was performed to assess the effect of mutant NFAT3 on proliferation of HaCaT cells. (d) An EGF-stimulated (20 ng/ml) soft agar assay was performed using HaCaT cells stably expressing wildtype NFAT3 or mutant NFAT3. Left panel shows representative photos. Right panel shows statistical analysis of colony number. (e) Representative photos and statistical analysis of 2-D colony formation of A431 stable cells expressing wildtype NFAT3 or mutant NFAT3. (f) Xenograft tumor growth of A431 stable cells expressing wildtype NFAT3 or mutant NFAT3 in nude mice. Points indicate mean values (n=6); bars indicate s.d. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001.