TNFRSF10A mediates ER stress-induced apoptosis in a DDIT3-dependent mechanism.
A, A549 cells were transfected with control (Ctrl) siRNA or TNFRSF10A siRNA #1 and #2. Forty-eight hours later, the cells were treated with DMSO, TG (1 μm), or Tm (1 μm) for another 24 h, and samples were analyzed by Western blot. B, A549 cells were transfected with control siRNA, DDIT3 siRNA, or TNFRSF10A siRNA #2 or co-transfected with DDIT3 and TNFRSF10A siRNA. After 48 h, the cells were treated with DMSO, TG (1 μm), or Tm (1 μm) for another 24 h, and samples were analyzed by Western blot using antibodies against DDIT3, TNFRSF10A, CASP8, CASP3, PARP1, and ACTB. C, A549 cells were transfected with Control siRNA, DDIT3 siRNA, or TNFRSF10A siRNA #2 or co-transfected with DDIT3 and TNFRSF10A siRNA. After 48 h, the cells were treated with DMSO, TG (1 μm), or Tm (1 μm) for another 24 h, and then cells were stained with annexin V-PE/7-AAD, and apoptosis was detected by flow cytometry analysis. Columns, mean of triplicate treatments; bars, ± S.D. The statistical differences between the two treatments were analyzed by two-sided unpaired Student's t tests (*, p < 0.05; **, p < 0.01).