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. 2013 Apr 22;288(24):17725–17733. doi: 10.1074/jbc.M113.453548

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — Attenuated NKp80-mediated cytotoxicity can be enhanced by overexpression of Syk. NK-92MI-NKp80, NK-92MI-NKp80/DGY, and NK-92MI-NKp80/ERF stably expressing high levels of wild type Syk were generated by transduction and assayed (A) for Syk co-immunoprecipitation with NKp80 and (B) for NKp80-mediated cytotoxicity in a redirected cytolysis assay. A, Syk-transduced NK-92MI-NKp80, NK-92MI-NKp80/DGY, and NK-92MI-NKp80/ERF cells were treated with pervanadate (2 min) and subsequently lysed. NKp80 was precipitated with anti-NKp80 mAb 5D12 and precipitates assessed for NKp80 tyrosine phosphorylation and co-precipitated Syk. NKp80 precipitation was monitored with a polyclonal anti-NKp80 reagent. For control, lysates were directly assessed for Syk. Note that amounts of NKp80 in lysates were below detection threshold (B) NK-92MI-NKp80, NK-92MI-NKp80-Syk, NK-92MI-NKp80/DGY-Syk, and NK-92MI-NKp80/ERF-Syk were co-cultivated with ⁵¹Cr-labeled P815 cells in the presence of anti-NKp80 mAb 5D12 and % cytolysis determined after 4 h of co-culture.