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Fig. 2 | Cell Communication and Signaling

Fig. 2

From: Receptor-interacting protein kinase 1 is a key mediator in TLR3 ligand and Smac mimetic-induced cell death and suppresses TLR3 ligand-promoted invasion in cholangiocarcinoma

Fig. 2

Smac mimetic sensitizes CCA cell lines to TLR3 ligand, Poly(I:C)-induced caspase-8 activation and apoptosis. a MMNK1, KKU100, and KKU213 cells were pretreated with Smac mimetic (50 nM MMNK1 and KKU100, and 5 nM KKU213) (Smac) or Smac mimetic and 20 μM zVAD-fmk (SZ) for 2 h. After that the cells were transfected with 2.5 μg/ml Poly(I:C) (PS, PSZ) for 24 h. TNF-α at 10 ng/ml and Smac mimetic or zVAD-fmk at the same concentration as with Poly(I:C) (TS, TSZ) were used as a positive control. Cell death was determined by Annexin V and propidium iodide staining followed by flow cytometry. Data from three independent experiments was presented as mean ± S.D.; * p < 0.05, **p < 0.01, *** p < 0.001 b KKU100 and KKU213 cells were treated as in (a) for 6 h and 12 h. Cell lysates were collected, after that the activation of caspase-8 and caspase-3 and cleavage of PARP-1 were analyzed by Western blot analysis. β-actin was served as loading control. Data shown was a representative of two independent experiments

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