Fig. 5

Verification of the proximal enhancer of the chicken NANOG (cNANOG) gene in chicken embryonic stem cells (ESCs). a-b Schematic diagram of the constructed cNANOG promoter vectors and luciferase activity in ESCs (a) and DF-1 cells (b). c Prediction of transcription factor (TF)-binding sites in the cNANOG promoter region from − 442 to − 250 bp. d Mutation analysis of putative CCAAT/enhancer-binding protein (CEBP)-binding sites in ESCs. e Luciferase activity of the − 442/+ 70 bp cNANOG promoter fragment compared with that of the mutated promoter. Significant differences are indicated as ns (no significance) and *** P < 0.001. Error bar represent the SEs for five replicate reactions