Human N-Myc Interactor (Nmi), an interferon (IFN)-induced protein, participates in downstream JAK-STAT pathway and inhibits proliferation of cancer cells．Nmi can bind Interferon-Induced Protein 35 (IFP35) and protect the latter from degradation．Previous research showed that these two proteins associated through their C-terminal domains．In this study, we constructed full length clones and N-terminal truncates of both Nmi and IFP35．Co-expression and GST pull down experiments showed that N-terminal domains of Nmi and IFP 35 are respectively sufficient for their association．We overexpressed and purified recombinant protein complexes afterward．Through immune fluorescence, we found that Nmi and IFP35 co-localized in nucleus in RAW264.7 cells．We also noticed that Nmi localized in nucleus and enriched on surface in normal 293A cells．Although Nmi granularly aggregated in cytoplasm after 24 h induction by interferon as reported, it came back to nucleolus by adding low concentration hydrogen peroxide (H₂O₂)．That result indicated that subcellular localization of Nmi can be affected by the oxidation environment of cells.