Figure 6. Mechanism of OMCP-mutIL-2 Signalling in NKG2D-Expressing Cells.

(a) STAT5 phosphorylation in isolated NK cells from splenocytes of A/J (left) or C57BL/6 mice (right) by increasing doses of cytokine. (b) Canonical IL-2 and NKG2D signaling pathways. (c) Jak1 phosphorylation of Ky1.1 NK cell line in vitro (d) Vav phosphorylation in freshly isolated splenic C57BL/6 NK cells. Representative of two separate experiments with full blots demonstrated in Supplementary Fig. 7. (e) Construction and phenotype of BWZ.36 LacZ reporter cell line (top) and expression of NKG2D (bottom left) and CD132 (bottom right) with parental cell shown in black, functional NKG2D/DAP12 WT in green and NKG2D/mutant Y2F DAP12 in blue. Phenotype is representative of two separate experiments. (f) β-galactosidase expression by BWZ.36 cell line bearing functional NKG2D/DAP12 (left panel), NKG2D/mutant Y2F DAP12 (middle), or parental BWZ. 36 cell line not expressing NKG2D/DAP12. Data are representative of three separate experiments with comparison performed by ANOVA for multiple comparisons or unpaired t-test between individual groups as indicated by the lines above the graphs.