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Fig. 2
Plasmid pEGFP-N1 has core sequences regulating the expression of isg15. A BLASTN alignment of pEGFP-N1 plasmid and isg15 gene sequences (including 2 kbp promoter upstream of TSS) with Kablammo (http://kablammo.wasmuthlab.org/). B Comparison of c-ori, c-CMV and c-HSV (DNAMAN) sequences showing a common 7 bp element, 5ʹ-GTTTGTT-3ʹ. C qRT-PCR results showing that the three mutated fragments of c-ori, c-CMV and c-HSV have weakened activation of isg15 expression in 3 hpf embryos compared with the unmutated fragments. D Responses of HEK 293T cells to transfection with the three DNA fragments and their corresponding mutated sequences. Cells’ responses were significantly weakened after the three sequence mutations. E, F Semi-quantitative PCR and qRT-PCR results showing that the methylation level of H3K4me3 in the isg15 promoter region was significantly increased. MW: DL5000 DNA Molecular Weight Marker