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. 2016 Aug 2;17(8):1246. doi: 10.3390/ijms17081246

Figure 5.

Figure 5

Promoter mutation assay for the transcription factor-binding sites in the nucleotide −320 to −240 region of the hST8Sia VI gene in physcion-induced SK-N-BE(2)-C cells. (A) Nucleotide sequences of the promoter region from −320 to −240 are shown; (B) After co-transfection of each construct into SK-N-BE(2)-C cells with pRL-TK, relative firefly luciferase activity was assessed. The Renilla luciferase activity derived from pRL-TK was used to normalize all firefly luciferase activity. Data are the means ± SD of triplicate measurements and are representative of three independent experiments. The circle and square indicate NF-Y and Pax-5 binding sites, respectively. The white and black colors indicate wild-type and mutant forms of these sites, respectively; (C) ChIP assay was conducted with primers amplifying the −379 and −131 region of the hST8Sia VI promoter and DNA isolated from chromatin immunoprecipitated with either anti-Pax-5 antibody or control IgG from SK-N-BE(2)-C cells treated for 24 h with or without 40 μM physcion.