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. 2017 Apr 13;12(4):e0175435. doi: 10.1371/journal.pone.0175435

Fig 8. Effects of DEX on interaction between NeuroD1 and E-box on Pomc promoter in AtT20 cells.

Fig 8

(A) Effect of of DEX on interaction between NeuroD1 and E-box on Pomc promoter examined by ChIP assay using E-box primer. ChIP assay was performed using digested chromatin extracted from the cells cultured in the presence (100 nM) or absence (control) of DEX for 30 min (B), 60 min (C), or 24 hrs (D). Chromatin fragments were immunoprecipitated either by normal rabbit IgG (negative control) or NeuroD1 antibody. Purified DNA was analyzed by qPCR using primers specific for E-box containing sequence on Pomc promoter. The expected size of E-box is 196 bp. Few qPCR products observed in the input samples were detected in the immunoprecipitation using normal IgG. Immunoprecipitated DNA was quantified by qPCR and normalized to the values obtained after amplification of unprecipitated 1% input DNA. Each point represents mean ± SEM (n = 3). **P<0.01, significantly different from the level of control group. These experiments were repeated three times with separate batches of cell preparation with consistent results.