Extended Data Fig. 4 |. TRAF2 and cFLIP interact, stimulate NF-κB signaling, and increase resistance of AML cells to the killing of cytotoxic T cells.

a, Relative NF-κB activities in 293T cells co-transfected with NF-κB reporter plus empty vector, p22-FLIP, p43-CFLIP, or full-length cFLIP (n=3 independent experiments). b, Relative NF-κB activities in 293T cells co-transfected with NF-κB reporter plus empty vector or tet-on cFLIP in the presence of dox (n=3 independent experiments). c, Co-immunoprecipitation assay of exogenous expressed FLAG-cFLIP and HA-TRAF2 in 293T cells. d and e, Overexpression of TRAF2 and cFLIP increase the resistance of monocytic AML cells to cytotoxic T cells. CFSE-stained THP-1 cells that overexpress TRAF2 or empty vector (EV) (d) or cFLIP or empty vector (e) were co-cultured with activated T cells at the different ratios for 12 hours and cell death was quantified. Left: Plots of percentage of dead cells versus E:T ration. Right: FACS analyses with E:T ratio of 2 (n=3 independent experiments). f, West blotting of pMLKL (pS358) and MLKL in THP-1 cells treated with coated IgG or anti-LILRB3 for 12 hours. g, Percentages of dead cells in THP-1 cells treated with anti-LILRB3 antibody or IgG in the presence of DMSO or NF-κB inhibitor QNZ (10 μM) (n=3 independent cell cultures). The data are presented as mean ± s.e.m, and p values were calculated by two-tailed t-test.