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Fig. 1. Synthesis and intracellular retention of IFN{gamma}. (A) WISH cells, untreated (lanes 1 and 2) or those transduced for two days with an empty vector control (lanes 3 and 4) or a vector expressing non-secreted IFN{gamma} (lanes 5 and 6) or a vector expressing non-secreted IFN{gamma} mutated in the NLS (lanes 7 and 8) were used. Proteins from cell extracts (odd-numbered lanes) or supernatants (even-numbered lanes) were separated by SDS-PAGE and probed with an antibody to IFN{gamma}. Detection was carried out by using chemiluminescence. (B) Quantitation of IFN{gamma} produced in L929 cells by ELISA. Cell extracts (odd numbers) and supernatants (even numbers) from L929 cells, transduced for two days with the empty adenoviral vector (column 1 and 2) or vector expressing non-secreted IFN{gamma} (column 3 and 4) or vector expressing non-secreted IFN{gamma} mutated in the NLS (column 5 and 6) were assayed for IFN{gamma} by ELISA.