Fig. 3: Cullin-1 inhibits TRAIL-induced apoptosis in HCT116 cells.
From: The SCFSkp2 ubiquitin ligase complex modulates TRAIL-R2-induced apoptosis by regulating FLIP(L)
a Western blot analysis of TRAIL-R2 DISC IP in HCT116 BAX-null cells transfected for 48 h with 30 nM control (SC) or a pool of Cullin-1 targeted (siCul1) siRNAs prior to treatment with zVAD-fmk (20 µM) for 2 h and then activation of the TRAIL-R2 receptor for 1 and 3 h. b Western blot analyses of apoptosis proteins in HCT116 cells transfected with 10 nM control (SC) or Cullin-1-targeted (siCul1) siRNAs for 48 h prior to treatment with rTRAIL (10 ng/mL) for 6 h. High content microscopy analysis of cell death in (c) HCT116 cells and (d) A549 and HT29 cells transfected with 10 nM control (SC) or Cullin-1-targeted (siCul1) siRNAs for 24 h prior to treatment with rTRAIL for an additional 24 h. A student’s T test was performed between SC and siCul1 samples (Error bars = SEM, ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ns = p > 0.05). e Western blot analysis of TRAIL-R2 DISC IP in HCT116 BAX-null cells transfected with 10 nM of a pool of FLIP(L)-specific or a pool of pan-FLIP siRNAs and treated with 10 µM zVAD-fmk for 16 h prior to DISC stimulation for 1 h.
