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. 2012 Feb 13;287(15):12405–12416. doi: 10.1074/jbc.M111.304469

FIGURE 4.

FIGURE 4.

NMNAT-1 and PARP-1 bind to promoter regions of commonly regulated target genes. A, ChIP-qPCR analysis of NMNAT-1 (FLAG) and PARP-1 at the promoter (P) and an upstream region (U; ∼10 kb from the transcription start site) of target genes in MCF-7 cells. B, ChIP-Western analysis of interactions between PARP-1 and NMNAT-1. MCF-7 cells expressing GFP or FLAG-NMNAT-1 (F-NMNAT-1) were used for PARP-1 and FLAG ChIP. The immunoprecipitated (IP) material was subjected to Western blotting for FLAG and PARP-1, respectively. The blots shown are representative of three independent experiments. C, GST-NMNAT-1 interaction assay with native PARP-1 from a nuclear extract. PARP-1 interaction with GST-NMNAT-1 bound to glutathione-agarose resin was detected by Western blot analysis. The blot shown is representative of three independent experiments. D, FLAG-based ChIP-qPCR analysis of NMNAT-1 localization to target gene promoters in control or PARP-1 knockdown (KD) cells. Each bar shown in A and D represents the mean of three or more independent experiments, and the error bars represent S.E.