Fig. 6
GMEB1 inhibited the formation of DISC upon TRAIL activation. a A549 cells and H1299 cells were seeded in 12-well plates. GMEB1 siRNA was transfected for 24 h and a non-sense siRNA was transfected as negative control. Cells were then treated with TRAIL with different concentrations for 6 h and harvested for western blot analysis. b A549 cells were seeded in 96-well plates. GMEB1 siRNA was transfected for 48 h and a non-sense siRNA was transfected as negative control. Cells were then treated with TRAIL [40 ng/ml] for 6 h and prepared for CASP8 activity detection. c A549 cells were seeded in 6-well plates. GMEB1 siRNA and FLAG-CFLARL plasmid were co-transfected for 48 h. Cells were then treated with TRAIL [40 ng/ml] for 6 h and prepared for CASP8 activity detection. d, e A549 cells were seeded in 96-well plates. GMEB1 siRNA was transfected for 48 h and a non-sense siRNA was transfected as negative control. Cells were then treated with TRAIL at different concentrations for 4 h and harvested for IP assay using FADD or CASP8 antibody