Figure 6: USP6 promotes NK activation in vivo, and directly induces NK activation in vitro.

A) Digested USP6/A673 tumors were subjected to flow cytometry. The percent of cells positive for NK1.1⁺ relative to total live cells in the digested tumor was quantified, as was CD69 and CD25 in the NK1.1⁺/CD45⁺ live population. See Supplemental Figures 12/13 for gating strategy. B) NK92 (effector, E) cells were co-incubated with USP6/RD-ES (GFP⁺) (target, T) cells at various effector E:T ratios overnight, in the absence or presence of dox. Percent survival of the USP6/RD-ES (relative to sample without NK92) was quantified by monitoring the GFP⁺/propidium iodide-excluded population. C/D) USP6/RDES and NK92 were co-cultured overnight with or without dox, then subjected to (C) flow cytometry to monitor surface CD69 and CD107a on the NK92 (CD45⁺/GFP⁻) cells, or (D) RT-qPCR to quantify IFNγ, CXCL9, and CXCL10 expression.