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, K^exp.
• 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 γ_c family members show measurable responses and EC₅₀ values.
• Fig. S20. Set of parameters for the extended γ_c family model.
• Table S1. List of antibodies used in this study.

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• 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