Supplementary Materials

Supplementary Materials for:

Intra- and Interdimeric Caspase-8 Self-Cleavage Controls Strength and Timing of CD95-Induced Apoptosis

Stefan M. Kallenberger, Joël Beaudouin, Juliane Claus, Carmen Fischer, Peter K. Sorger, Stefan Legewie, Roland Eils*

*Corresponding author. E-mail: r.eils@dkfz.de

This PDF file includes:

  • Text S1. Data analysis.
  • Text S2. Model implementation and fitting.
  • Text S3. Model analysis and verification.
  • Fig. S1. Procaspase-8 and its cleavage products.
  • Fig. S2. Total cleavage probe concentrations and apoptosis times.
  • Fig. S3. Single-cell caspase-8 trajectories for CD95-HeLa and wild-type HeLa cells.
  • Fig. S4. Single-cell trajectories for NES- and FRET-probe cleavage related to caspase-8 activity.
  • Fig. S5. Western blot of wild-type and prodomain mutant caspase-8.
  • Fig. S6. Model graphs for oligomerization-driven receptor activation and caspase-8 activation.
  • Fig. S7. Caspase-8 Western blot time series were used for model fitting of population observables.
  • Fig. S8. BID and tBID Western blot time series served for model fitting of population observables.
  • Fig. S9. Estimation of average FADD and p55 amounts by calibrated Western blot.
  • Fig. S10. Estimation of average BID amounts by calibrated Western blot.
  • Fig. S11. Estimation of average cleavage probe amounts by calibrated Western blot.
  • Fig. S12. Flow cytometry of cells immunostained for CD95.
  • Fig. S13. Flow cytometry of cells immunostained for p55, FADD, and BID.
  • Fig. S14. Fit qualities of extended cis/cis and cis/trans model variants.
  • Fig. S15. Reaction fluxes for the cis+trans/cis+trans topology in separate cleavage reactions.
  • Fig. S16. Reaction fluxes and half-lives of caspase-8 intermediates for the cis/trans topology.
  • Fig. S17. Reaction fluxes and half-lives of caspase-8 intermediates for the trans/trans topology.
  • Fig. S18. Monte Carlo modeling supports discrimination between cis/cis and cis/trans topologies.
  • Fig. S19. Predicted coefficients of variation of apoptosis times at different CD95 ligand concentrations.
  • Fig. S20. Variability of active receptor fractions for CD95-HeLa and wild-type HeLa cells.
  • Fig. S21. Model predictions of the effects of twofold increases in initial protein concentrations.
  • Table S1. Reactions of the complete model cis+trans/cis+trans and smaller variants.
  • Table S2. Reaction rates in the complete cis+trans/cis+trans model and smaller variants.
  • Table S3. Model equations of variants cis+trans/cis+trans and cis/trans.
  • Table S4. Estimates of protein numbers in CD95R-HeLa and wild-type HeLa cells.
  • Table S5. Receptor number estimates.
  • Table S6. Initial values and lower and upper limits for estimations of initial concentration.
  • Table S7. Cellular and cytosolic volumes, cell surface areas, and average fluorescence intensities.
  • Table S8. Parameters of the fitted cis/trans model.
  • References (4961)

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Other Supplementary Material for this manuscript includes the following:

  • Caspase-8 model MatLab scripts (zip file of .m files)
  • Caspase-8 model SBML files (zip file of .xml files)
  • Caspase-8 single-cell data (Excel)

[Download Caspase-8 model MatLab scripts]

[Download Caspase-8 model SBML files]

[Download Caspase-8-cell data]


Citation: S. M. Kallenberger, J. Beaudouin, J. Claus, C. Fischer, P. K. Sorger, S. Legewie, R. Eils, Intra- and Interdimeric Caspase-8 Self-Cleavage Controls Strength and Timing of CD95-Induced Apoptosis. Sci. Signal. 7, ra23 (2014).

© 2014 American Association for the Advancement of Science