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Assay for NF-κB binding to ATM promoter.

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posted on 20.02.2013, 11:49 by Xiaoqian Ma, Lifang Yang, Lanbo Xiao, Min Tang, Liyu Liu, Zijian Li, Mengyao Deng, Lunquan Sun, Ya Cao

A, Biotin-labeled wild type ATM-NF1 was incubated with the nuclear extracts from CNE1 (lane 2), CNE1-LMP1 (lanes 3–6) or CNE1-LMP1 treated with Dz1 (lane 7) and CNE1-LMP1 treated with oligonucleotide control ODN (lane 8). A 200-fold excess of noncompetitive unlabelled Stat3 oligonucleotide (U, lane 4), unlabeled mutated ATM-NF1 (mut, lane 5) and unlabeled wild-type ATM-NF1 (wt, lane 6) were included in the respective reactions. Oct1 bands served as loading control. B, Biotin-labeled wild type ATM-NF1 was incubated with the nuclear extracts from CNE1-LMP1 treated with antibodies to p65 (lane 3), p50 (lane 4) and p52 (lane 5). C, Biotin-labeled wild type ATM-NF2 was incubated with the nuclear extracts from CNE1 (lane 2), CNE1-LMP1 (lanes 3) or CNE1-LMP1 treated with Dz1 (lane 4) and CNE1-LMP1 treated with oligonucleotide control ODN (lane 5). Unlabeled wild-type ATM-NF2 (wt, lane 6) and unlabeled mutated ATM-NF2 (mut, lane 7) were included in the respective reactions. Oct1 bands served as loading control. D, Biotin-labeled wild type ATM-NF2 was incubated with the nuclear extracts from CNE1-LMP1 treated with antibodies to p65 (lane 3), p50 (lane 4) and p52 (lane 5).Oct1 bands served as loading control.

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