TNF-α and IFN-γ cooperate in the activation of macrophages. TNF-α-dependent activation of NF-κB is stronger in the presence of IFN-γ. STAT-1α associates with TNFR1 in TNF-α-treated cells, and this association attenuates TNF-α-mediated NF-κB activation. We hypothesized that nuclear localization of STAT-1α due to IFN-γ signaling would preclude it from being recruited to the TNFR1 and therefore enhance TNF-α-induced NF-κB activation. In the RAW264.7 macrophage cell line, TNF-α treatment indeed recruits STAT-1α to the TNFR1, and this association is abrogated when cells are exposed to IFN-γ. TNF-α treatment induces a more robust activation of NF-κB in STAT-1α-deficient cells, and restoration of STAT-1α inhibits TNF-α-dependent NF-κB activation. Our results suggest that a receptor-proximal level of cross-talk exists between these two cytokine pathways: IFN-γ limits STAT-1α availability to the TNFR1 by depleting STAT-1α from the cytoplasm, thus allowing for optimal NF-κB activation upon TNF-α ligation.