Nuclear localization of STAT1 in TO cells after stimulation with IFN-a1 and IFNγ. The cells were treated with IFN-a1 (10 U/mL) for 45 or 90 min or IFNγ (200 ng/mL) for 45 min or left unstimulated. Nuclear extracts (N) and cytoplasmic fractions (C) were separated using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo scientific). The extracts were subjected to SDS-PAGE and Western blotting. STAT1 was detected in both N and C fractions, but lesser STAT1 was observed in the nuclear fraction of the unstimulated cells. An antibody directed against a strictly lysosomal protein, Cathepsin D (CatD) was used as a control for cytoplasmic localization. After stripping the membrane, an actin antibody was applied as a loading control. M = MagicMark molecular weight marker.