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. 2015 Dec 14;291(5):2379–2388. doi: 10.1074/jbc.M115.685008

FIGURE 2.

FIGURE 2.

TRAIL induces robust up-regulation of CAS in a caspase-8-dependent manner and independently of MOMP. a, MCF10A cells were treated with 30 ng/ml TRAIL and harvested for Western blot analysis at the indicated time points. b, MCF10A cells were pretreated for 30 min with 20 μm of a pan-caspase inhibitor (Z-VAD-fmk), a caspase-3 inhibitor (C3i, DEVD-fmk), or a caspase-8 inhibitor (C8i, IETD-fmk), followed by 40 ng/ml TRAIL for 3 h, and subsequently harvested for Western blot analysis. Right panel, quantification of CAS levels (mean ± S.D.) relative to untreated cells from three independent experiments. c, MCF10A cells stably expressing control shRNA or one of two different shRNAs against caspase-8 were treated with 30 ng/ml TRAIL for 3 h, and expression of the indicated proteins was analyzed by Western blot. The asterisk indicates a nonspecific band. d, WT and Bcl-xL-overexpressing MCF10A cells stably expressing SMAC-cherry were treated with 40 ng/ml TRAIL for 2.5 h, and imaged for the onset of MOMP. e, WT and Bcl-xL-overexpressing MCF10A cells were treated with 40 ng/ml TRAIL for 3 h and harvested for Western blot analysis with the indicated antibodies.