Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Soluble factors released by activated cytotoxic T lymphocytes interfere with death receptor pathways in neuroblastoma

doi: 10.1007/s00262-007-0412-2

Figure Lengend Snippet: Supernatant from activated CTLs both induce caspase-8 expression and modulate its activity in NB cells. a NB cell lines were treated for 48 h with either medium alone, or in the presence of 10% v/v of control supernatant (CS) or activated supernatant (AS). The expression of caspase-8 in total cell lystaes was monitored by Western blot using a specific antibody recognizing the 55/50 kD pro-form of caspase-8 as well as the 40/36 and 23 kD cleavage products obtained upon activation. Expression of actin was used as a control of sample loading. b The activity of caspase-8 in NB cells propagated for 24, 48 and 72 h in complete medium alone (thin open histograms), or in the presence of either CS (filled histograms) or AS (bold open histograms) was measured by flow cytometry using fluorescent caspase-8 specific substrates as described in Materials and Methods. Numbers show percentage of AS-treated cells positive for active caspase-8

Article Snippet: The following antibodies and reagents were used for Western blot analysis: the mouse monoclonal antibody specific to β-actin was purchased from Sigma Chemical Co (St Louis, USA) and the mouse anti-human caspase-8 mAb was from BD Pharmingen.

Techniques: Expressing, Activity Assay, Western Blot, Activation Assay, Flow Cytometry