Dear Editor,

We analysed in more detail all cleavage products of procaspase-8 in the DISC formed upon stimulation of the B-lymphoblastoid cell line SKW6.4. Cells were treated with LZ-CD95L for different time points, followed by immunoprecipitation of the DISC components with the agonistic anti-APO-1 and by immunoblotting with a panel of mAbs. The epitopes recognised by these antibodies are shown in Figure 1a. In addition to the established components of the DISC, including full-length procaspase-8 (p55/p53) and its cleavage product p43/p41, we clearly detected the p18 and p10 subunits of procaspase-8 in the DISC (Figure 1b). The presence of p10 and p18 at the DISC was also observed using other T-cell lines Hut78, CEM, H9 as well as primary T cells (data not shown). The detection of these products became possible because we used different conditions for electrophoresis, focusing on lower molecular weight products in comparison to previous studies. In addition, the presence of the p10 subunit was detected using the C5 antibody that had not been used in the analysis of the lower molecular weight procaspase-8 subunits in the CD95 DISC before.

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