Fig. 5

SLC39A7 transporter function is required to relieve ER stress, restore TNFR1 surface expression and induce cell death. a KBM7 FADD- SLC39A7- cells lentivirally transduced with either V5-tagged SLC39A7 or GFP were lysed and immunoblotted with the indicated antibodies, KBM7 FADD- sgRen cells serve as control. Immunoblots shown are representative of two independent experiments, asterisk (*) indicates non-specific band. b Flow cytometry analysis for TNFR1 surface expression in KBM7 FADD- SLC39A7- cells reconstituted with either V5-tagged SLC39A7 or GFP. KBM7 FADD- sgRen and empty KBM7 FADD- SLC39A7- cells serve as positive and negative control, respectively. Data shown are representative of two independent experiments. c Cell viability in KBM7 FADD- SLC39A7- cells reconstituted with either V5-tagged SLC39A7 or GFP, KBM7 FADD- sgRen and empty KBM7 FADD- SLC39A7- serve as controls. Cells were treated overnight (16 h) with TNFα (10 ng/ml), SMAC mimetic (0.5 µM), or a combination thereof. Cell viability was assessed using a luminescence-based readout for ATP (CellTiter Glo). Data represent mean value ± s.d. of two independent experiments performed in triplicates. d Cell viability in KBM7 FADD- SLC39A7- cells stably reconstituted with GFP or the indicated V5-tagged SLC39A7 constructs. Cells were treated as indicated for 24 h and cell viability was assessed as in c. Data represent mean value ± s.d. of two independent experiments performed in triplicates. e KBM7 FADD- SLC39A7- or KBM7 FADD- cells stably reconstituted with the specified constructs were lysed and subjected to immunoblotting with the indicated antibodies. Asterisk (*) indicates non-specific band