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. Author manuscript; available in PMC: 2014 Jul 15.
Published in final edited form as: Exp Cell Res. 2013 Apr 12;319(12):1902–1911. doi: 10.1016/j.yexcr.2013.04.005

Fig. 5. Effects of ODC overexpression and putrescine on MAT2A promoter activity.

Fig. 5

(A) RKO cells were transfected transiently MAT2A promoter construct −571/+60-LUC or pGL-3 enhancer vector and co-transfected with ODC overexpression vector or treated with putrescine (100 pM) for 24 hours. Results represent mean±SEM from 3 independent experiments performed in triplicate. Data are expressed as relative luciferase activity to that of pGL-3 enhancer vector control. *p<0.001 vs. pGL-3, †p<0.002 vs. −571/+60-LUC construct control. (B) Cells were treated with putrescine (100 pM) for 24 hours and processed for ChIP using an anti-c-Jun antibody and a ChIP assay kit according to the manufacturer’s protocol. The AP-1 element in MAT2A promoter fragment from c-Jun immunoprecipitates was amplified by PCR using the corresponding primers. The regulatory fragment was directly amplified by PCR of genomic DNA from each group as input. Value are average of 3 separate experiments ± SEM. *p<0.001 vs. untreated control.