Figure 4

ChIP assay to show that NFAT proteins are associated with human TMP21 promoter. ChIP assay was performed as described in the Material and Methods. The chromatin was isolated HEK293 (A) or SH-SY5Y cells (B), and then sheared from the cells treated with cross-link reagent formaldehyde. For isolation of NFAT binding complex, the chromatin solution was incubated with NFAT antibody (Santa Cruz). PCR amplifications were performed using TMP21 promoter-specific primers, with samples from the sheared chromatin alone (lane 2), non-NFAT antibody control (lane 3) and NFAT-immunoprecipitating chromatin sample (lane 4). β-actin was used as internal control.