Nfatc2

ChIP assay was done with the ChIP Assay kit (Millipore) following the vendor’s instruction. In brief, cells were cross-linked with 1% formaldehyde at 37 °C for 10 min, followed by incubation with 150 mM glycine for 5 min at room temperature to stop cross-linking. The cells were then harvested in SDS lysis buffer containing protease inhibitors. After the lysates were sonicated to shear the DNA into fragments of 200–1000 bps, immunoprecipitation with NFATc2 (Abcam) or nonspecific IgG was carried out overnight at 4 °C. Protein G Sepharose (GE Healthcare, Little Chalfont, Buckinghamshire, UK) was added to the suspension to collect the antibody/NFATc2/DNA complex, followed by rinsing with each of Low Salt Immune Complex Wash Buffer, High Salt Immune Complex Wash Buffer, LiCl Immune Complex Wash Buffer, and TE Buffer. The precipitated products were then eluted with elution buffer containing 1% SDS, 0.1 mol/L NaHCO₃, reversed the NFATc2/DNA cross-link with 5 mol/L NaCl at 65°C for 4 h. The DNA was finally eluted, and purified by PCR Purification Kit from Qiagen. PCR was performed with primers listed in Supplementary Table 8.