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Figure 8 Detection of promoter activities of the 5′-flanking region of chicken GPR12 in cultured chicken fibroblast cells line (DF-1). Various lengths of the 5′-flanking regions of cGPR12 were cloned into pGL3-Basic vector for the generation of multiple promoter-luciferase constructs (−3050/+277 Luc and −1962/+277 Luc). These promoter-luciferase constructs were then co-transfected into DF-1 cells along with pRL-TK vector, and their promoter activities were determined by dual-luciferase reporter assays. The transcriptional start site identified by 5′-RACE was designated as “+1” (see Figure S1). Each value represents the mean ± SEM of four replicates (N = 4). *** p < 0.001 vs. pGL3-Basic vector.