FIGURE 5.

NMNAT-1 regulates PARP-1 activity at target gene promoters. ChIP-qPCR and RT-qPCR analyses were carried out using MCF-7 cells ectopically expressing GFP, FLAG-NMNAT-1 (F-NMNAT-1) wild type (NWT), or FLAG-NMNAT-1 W169A enzymatic mutant (NMA), as indicated by shading as shown in A. For all panels, the bars shown represent the mean of three or more independent experiments, and the error bars represent S.E. A–C, ChIP for NMNAT-1 (FLAG) (A), PARP-1 (B), and PAR (using the PAR monoclonal antibody clone 10H) (C) at the promoter (P) and an upstream region (U; ∼10 kb from the transcription start site) of target genes. D, RT-qPCR analysis of target gene expression. β-Actin mRNA was used as an internal reference for data normalization. The results are presented as expression levels relative to those observed in the GFP control cells. Statistical significance was determined by two-tailed Student's t test. *, p value <0.05.