Enhanced NKR-P1C-induced signaling in MIST-deficient NK cells. (A) NKR-P1C-induced intracellular calcium flux in MIST-deficient NK cells. Wild-type (+/+) and MIST-deficient (-/-) NK cells loaded with Indo 1-AM were stimulated with anti-NK1.1 antibody. The ratio of fluorescence detected in FL5-FL4 was monitored by flow cytometry. Acquisition was interrupted once to add cross-linking goat F(ab′)2 antibody (arrows) and again to add 1 μg/mL ioomycin (arrowheads). (B) Enhanced phosphorylation of PLCγ2, ERK, and p38 MAP kinases in MIST-deficient NK cells upon NKR-P1C stimulation. Cell lysates from wild-type (+/+) and MIST-deficient (-/-) NK cells stimulated with anti-NK1.1 antibody were immunoblotted with the indicated antibodies. Figures give induction levels over zero time points, determined by densitometry after normalizing for loading.
