FIGURE 4.

C/EBPβ binds multiple sites on the RCAN1–4 promoter. A, a schematic of the location of the various templates used for EMSA experiments is shown. B and C, EMSAs were carried out using template 2, and nuclear extracts from C2C12 myoblasts were transfected with LAP, Cn, and NFATc1 expression vectors. 40-fold excess unlabeled template containing a consensus binding site specific for C/EBPβ (B) or for NFAT (C) was used to compete for binding. D, gradients of excess cold C/EBPβ, NFAT, and nonspecific (non-spe) template (ranging from 20 to 500-fold) were used to compete for binding to template 2. E, EMSA analysis was carried out using fragments 1, 2, 3, and 4 as templates.