Figure 5.

Sck binding is required for CD95-induced PI3K/MAPK cascades. (a) Phosphorylation of AKT and ERK upon treatment with the indicated doses of CD95L-T4 for the different time points is shown in PanD24 cells. (b) Phosphorylation of AKT and ERK upon treatment with the indicated doses of CD95L-T4 for the different time points is shown in PANC-1 cells. (c) TranSignal SH2 domain arrays showing binding of endogenous CD95 to the SH2 domain of Sck in PANC-1 cells. (d) Co-immunoprecipitation of CD95 and Sck is shown in PANC-1 cells. Increased recruitment of Sck is observed after 15 min stimulation (upper panels). CD95 was immunoprecipitated from PanD24 cells treated with CD95L-T4 for 5 or 15 min. The immunoprecipitates were immunoblotted with anti-Sck antibody (lower panels). Data are representative of two independent experiments. (e and f) PanD24 (e) and PANC-1 (f) cells were transiently transfected with either non-targeting siRNA (Scrm) or Sck-siRNA (Sck KD) upon treatment with the indicated doses of CD95L-T4 and different time points