Fig 2. Interference with the IFNGR pathway and SLFN11 protects HAP1 from T cell- and IFNγ-mediated toxicity.

A) Parental HAP1 or two IFNGR1 KO clones were exposed to T cells at the indicated effector: target ratio for 24 h. 7 days after T cell exposure, surviving cells were stained with crystal violet. B-C) HAP1 cells transduced with a Cas9-encoding lentiviral vector with either a non-targeting sgRNA (sgRNA ctrl) or two independent sgRNA targeting SLFN11 (sgRNA SLFN11#1 and#2) were exposed to either T cells at the indicated effector: target ratio for 24 h (B), or to IFN-γ at the indicated concentrations for the whole duration of the experiment (C). 7 days after T cell or IFN-γ exposure, surviving cells were stained with crystal violet. D) HAP1 cells transduced with a Cas9-encoding lentiviral vector with either a non-targeting sgRNA (sgRNA ctrl) or two independent sgRNA targeting SLFN11 (sgRNA SLFN11#1 and#2) were exposed to the indicated concentrations of IFN-γ. 48 hours after IFN-γ exposure, cell viability was assayed by analysis of metabolic activity. E) HAP1 cells transduced with a control lentiviral vector or with two independent lentiviral vectors encoding SLFN11-targeting shRNA were exposed to the indicated concentrations of IFN-γ. 48 hours after IFN-γ exposure, cell viability was assayed by analysis of metabolic activity. F) Validation of SLFN11 KD by western blot analysis of untreated or IFN-γ treated (10 ng/ml, 24 h) HAP1 cells. G) HAP1 cells that were either untreated or pre-incubated for 1 h with the indicated compounds (20 μM for necrostatin, 10 μM for Q-VD-OPh) were exposed to the indicated concentrations of IFN-γ. 48 hours after IFN-γ exposure, cell viability was assayed by analysis of metabolic activity. * p<0.05, ** p<0.01, *** p<0.001.