Figure 4.

Nutlin-3 treatment induces Fas-dependent apoptosis in TC cells. (a) Following the indicated treatment, TC cells were harvested and Fas membrane expression was determined by flow cytometry. Values are depicted as MFI. Values are the mean±S.D. of three experiments. (b) TC cells were treated as indicated and Fas-membrane expression has been determined by immunofluorescence, representative example of three independent experiments. Scale bar: 30 μM. (c) Downregulation of p53 reduces Fas membrane expression after Nutlin-3 treatment in wild-type p53-expressing TC cells, compared with control; values are the mean±S.D. of three experiments; ^#P<0.05; ^*P<0.01; ^**P<0.005. (d) Fas acts pro-apoptotic after 24 h Nutlin-3 treatment in TC cells. After successful downregulation of FasL or blocking of FasL with NOK-1, a decrease in PARP cleavage and active caspase-8 is observed in Tera and 2102EP; a representative example of three independent experiments is shown. (e) Decreased apoptotic response after blocking or suppression of FasL, in TC cells treated with Nutlin-3; values are the mean±S.D. of three experiments