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. 2010 Mar 17;285(21):15894–15905. doi: 10.1074/jbc.M109.098376

FIGURE 2.

FIGURE 2.

Lack of NF-E2 (p45) impairs the assembly of protein complexes at LCR HS2 and at the adult βmaj-globin gene promoter. A, ChIP analysis of protein chromatin interactions in LCR HS2 and the adult βmaj-globin gene promoter in MEL cells incubated with or without 1.5% DMSO for 3 days. After cross-linking MEL cells with 1% formaldehyde, chromatin was isolated, fragmented by sonication, and subjected to immunoprecipitation with antibodies against NF-E2 (p45), MafK, USF1, USF2, CBP, and TFIIB. Reactions with the IgG antibody served as a negative control. The DNA was purified from the precipitate and subjected to qPCR using primers specific for LCR HS2 and the adult βmaj-globin gene promoter as indicated. Error bars, S.E. of three independent experiments (**, sample versus IgG, p < 0.05; ΔΔ, sample versus IgG, 0.05 < p < 0.1; *, DMSO versus no DMSO, p < 0.05; Δ, DMSO versus no DMSO, 0.05 < p < 0.1). B, ChIP analysis of protein chromatin interactions in LCR HS2 and the adult βmaj-globin gene promoter in CB3 cells incubated with or without 1.5% DMSO for 3 days. DNA was isolated from immunoprecipitated material and analyzed as described in A. Error bars, S.E. of three independent experiments (symbols are as in A). C, comparative ChIP analysis of protein chromatin interactions in uninduced CB3, MEL, and CB3/NF-E2 cells. Cross-linked chromatin was precipitated with IgG or antibodies against NF-E2 (p45), USF1, or USF2, and DNA was analyzed as described in A. Error bars, S.E. of three independent experiments (*, CB3/NF-E2 versus CB3, p < 0.05; Δ, CB3/NF-E2 versus CB3, 0.05 < p < 0.1; **, CB3/NF-E2 versus MEL, p < 0.05; ΔΔ, CB3/NF-E2 versus MEL, 0.05 < p < 0.1; ***, CB3/NF-E2 versus CB3 and MEL p < 0.05).