Reduction in miR-132, -212, and -200a expression in IL-12 tolerized NK cells inversely correlated with IFN-γ production. (A) NK cells were cultured with (tolerized) or without (untolerized) 10 ng/mL IL-12 continuously for 24 hours in the presence of IL-2. Cells were then washed twice with PBS and cultured in complete growth medium with IL-2 for additional 0, 12, or 24 hours (IL-12 withdrawal). At each time point after IL-12 withdrawal, cells were challenged with 10 ng/mL of IL-12 for 8 hours in the presence of IL-2 before analysis of IFN-γ protein production in culture supernatant by ELISA. (B) qRT-PCR analysis of total small RNA for miR-132, miR-212 and miR-200a expression. NK cells were cultured as in panel A. The results are mean ± SEM of representative experiment with NK cells from 3 different donors. *P < .05, **P < .01 versus IL-12 untolerized cells.