Supplementary Materials

Supplementary Materials for:

Cell-to-Cell Variability Analysis Dissects the Plasticity of Signaling of Common γ Chain Cytokines in T Cells

Jesse W. Cotari, Guillaume Voisinne, Orly Even Dar, Volkan Karabacak, Grégoire Altan-Bonnet*

*Corresponding author. E-mail:

This PDF file includes:

  • Computing cytokine sensitivity from affinity constants
  • Maximum a posteriori estimation of the affinity constants
  • Model enumeration and selection
  • Error estimation
  • Fig. S1. Flow cytometry provides accurate measures of relative protein abundance.
  • Fig. S2. CCVA analysis of cell populations from Fig. 1.
  • Fig. S3. IL-7Rα abundance does not correlate with variation in IL-7 EC50.
  • Fig. S4. Responsiveness to TSLP demonstrates the signaling functionality of IL- 7Rα.
  • Fig. S5. Kinetics of the chemical equilibrium for the pSTAT5 response.
  • Fig. S6. The pSTAT5 response is linear with the number of bound IL-7 molecules per cell.
  • Fig. S7. Cytokine depletion is negligible and does not affect the IL-2 or IL-7 EC50 values.
  • Fig. S8. Methodology for absolute receptor quantification.
  • Fig. S9. Estimation of the set of "measured" affinity constants, Kexp.
  • Fig. S10. A nondirect influence of α2 on γ cannot reproduce the dependency of IL-7 sensitivity on α2.
  • Fig. S11. Indexing of all relevant models.
  • Fig. S12. The experimentally determined affinity constants cannot quantitatively account for the effect of IL-2Rα on IL-2 and IL-7 EC50 values.
  • Fig. S13. Fitting without constraints on the affinity constants leads to great divergence from their measured values.
  • Fig. S14. Determination of the optimal value of ρ as a parameter constraint for model fitting.
  • Fig. S15. Model selection with corrected AIC.
  • Fig. S16. Determination of the 95% uncertainty intervals for the parameters from the Profile Likelihood (PL).
  • Fig. S17. Two-dimensional sections of the posterior distribution for the equilibrium constants for ρ → ∞(unconstrained fit).
  • Fig. S18. Two-dimensional sections of the posterior distribution for the equilibrium constants for ρ = 1 (constrained fit).
  • Fig. S19. pSTAT5 dose responses for γc family members show measurable responses and EC50 values.
  • Fig. S20. Set of parameters for the extended γc family model.
  • Table S1. List of antibodies used in this study.

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Other Supplementary Material for this manuscript includes the following:

  • ScatterSlice software and instruction manual

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Citation: J. W. Cotari, G. Voisinne, O. E. Dar, V. Karabacak, G. Altan-Bonnet, Cell-to-Cell Variability Analysis Dissects the Plasticity of Signaling of Common γ Chain Cytokines in T Cells. Sci. Signal. 6, ra17 (2013).

© 2013 American Association for the Advancement of Science