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. 2014 Dec 16;9(12):e115249. doi: 10.1371/journal.pone.0115249

Figure 8. NFAT3 protein binds CXCR4 promoter.

Figure 8

HeyA8 cells were cultured as monolayer and as 3D spheroids (100,000 cells per spheroid) as described in materials and methods. (A) CXCR4 gene sequence according to GeneBank accession number AF005058. Regions with putative transcriptional factor binding consensus sequences are underlined and in bold. The block arrows indicate real-time PCR primers used for the chromatin immunoprecipitation analysis. (B) Chromatin immunoprecipitation of adherent HeyA8 cells and 3D spheroid showed significant (p <0.005) increase in relative abundance of NFAT3 and Pol II on the CXCR4 promoter. (C) Similar to 3D spheroids, adherent cells treated with 20 µM of U0126 showed significant (p <0.05) increases in relative abundance of NFAT3 and PolII on the CXCR4 promoter. Average values were derived from 2 independent IPs with qPCRs for each IP performed in duplicate (n = 4).