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Fig. 4. Phosphorylation of STAT1{alpha} by intracellular IFN{gamma} is independent of the IFN{gamma} C-terminal NLS. WISH cells, untreated (lane 1) or treated with an empty vector control (lanes 2), a vector expressing non-secreted IFN{gamma} (lanes 3) or a vector expressing non-secreted IFN{gamma} with a mutated NLS (lanes 4) were allowed to grow for 8 hours. Proteins from whole cell extracts were electrophoresed and probed with an antibody to phospho-STAT1 (upper panel). Filter was stripped and re-probed with an antibody to STAT1{alpha} (lower panel). Detection was by chemiluminescence.