Fig. 4

CPP32 and FLICE expression in Jurkat cells, in response to treatment with sFas and with VEINCTR-N. (Left graphics) Immunoblotting of cell extract shows the increased expression of both CPP32 (upper graphic) and FLICE (lower graphic) and the presence of cleaved CPP32. Rabbit antiserum to CPP32 detected the 12.0 kDa active subunit of that protease in cultures treated with both soluble forms of Fas. Mouse MoAb to FLICE also revealed in these cultures the proenzymatic form in amounts comparable to that induced by CH11. Control untreated cells showed defective expression of either native or activated enzymes. Numbers refer to molecular weight (m.w.) (Right graphics) — CPP32, FLICE, and ICE mRNA analysis by reverse transcription polymerase chain reaction (RT-PCR) in Jurkat cells incubated with either soluble Fas (sFas) or VEINCTR-N, compared with treatment with CH11 and tumor necrosis factor α (TNFα). Both CPP32 and FLICE mRNA were expressed variably in cells treated with these soluble forms, compared with the control untreated cells. The semiquantitative Fluor-S evaluation revealed an increase of relative trace quantity values in apparent concordance with those detected in the control cell preparations treated with either CH11 or TNFα. By contrast, ICE was only expressed by cells supplemented with TNFα, suggesting that suppression of Jurkat cells in cultures treated with either sFas or VEINCTR-N was mediated by their Fas-pathway activation. As expected, the β-actin expression as internal control, was not influenced. Numbers are bp size.