Quantification of experimental data of CD95 signaling.

(A) The experimental data on c-FLIP (red) and procaspase-8 ratios to FADD (blue), respectively, were taken from Schleich et al. [20]. The corresponding extrapolation of these ratios for HeLa-CD95 cells (black) describes the chain configuration dependency on the CD95L stimulation dose. (B) Western blot analysis of DISC immunoprecipitation (IP) from HeLa-CD95 cells stimulated with 250 ng/ml CD95L for indicated time intervals. See S3 Fig for more details. (C) Representative example of application of quantitative Western blot data for modeling. Western blot analysis of signaling in HeLa-CD95 cells stimulated as in (B). One out of three representative experiments is shown (Complete data set is presented in S4 Fig). The model predictions (blue) are compared to the experimentally measured values (red). Means and SDs are shown. (D) Representative example of imaging flow cytometry data application for modeling. HeLa-CD95 cells were stimulated with 250 ng/ml CD95L and stained with antibodies against NF-κB subunit p65 and active caspase-3. The nucleus was stained with 7AAD. The probability density function (PDF) of the similarity score of the p65 and nuclear signal as well as the intensity of the caspase-3 signal were analyzed (red) and compared to the model predictions (blue). * = nonspecific band.