This PDF file includes:

• section S1. Description of the spatial stochastic models and the simulations
  
• table S1. Measured IFN receptor diffusion constants under different conditions.
  
• table S2. Calculated binding probability p_on for different microscopic diffusion constant Dmicro.
  
• table S3. Parameter scenarios (D_R1/R2, Δt, r_on, and p_on) and mean lifetimes τ for fitting hop probabilities.
  
• table S4. Experimental properties of primary and SCs.
  
• fig. S1. Comparison of non-colocalization criteria.
  
• fig. S2. Effect of the binding probability on the simulated receptor complex lifetime.
  
• fig. S3. Simulation of ternary complex lifetime in the hierarchical plasma membrane model.
  
• fig. S4. Orthogonal labeling of IFNAR1 and IFNAR2 with monovalent QD.
  
• fig. S5. High-fidelity colocomotion analysis.
  
• fig. S6. Role of the MSK for receptor diffusion.
  
• fig. S7. Model of the stochastic MSK meshwork used for simulations.
  
• fig. S8. Fitting of hop probabilities of the MSK model.
  
• fig. S9. Fitting of association probabilities.
  
• fig. S10. Sample simulations of ternary complex dissociation and reassociation.
  
• fig. S11. Dependency of simulated receptor complex lifetime on MSK hop probability.
  
• fig. S12. Effect of the reaction radius on the simulated receptor complex lifetime.
  
• fig. S13. TALM and pcTALM analyses of receptor confinement.
  
• fig. S14. Primary and secondary confinement confirmed by tracking receptors at 2000 Hz.
  
• fig. S15. Compartment size and dwell time determined by DBSCAN analysis of fast TALM images.
  
• fig. S16. Spatial stochastic model for simulating hierarchical confinement zones.
  
• fig. S17. Fitting of hop probabilities in the refined MSK model.
  
• fig. S18. Experimental trajectory length histograms for different colocalization cutoffs.
  
• fig. S19. Channel alignment for colocalization analysis.
  
• fig. S20. Determination of the observation window for time-lapse pcTALM.
  
• Legends for movies S1 to S7
  
• References (69–71)

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Other Supplementary Material for this manuscript includes the following:

• movie S1 (.avi format). Controls confirming the labeling specificity.
  
• movie S2 (.avi format). Dual-color single-molecule imaging of QD-labeled IFNAR1 and IFNAR2 in a live cell.
  
• movie S3 (.avi format). Characterization of receptor diffusion before and after IFN stimulation.
  
• movie S4 (.avi format). Assembly and dissociation of ternary complexes revealed by single-molecule tracking.
  
• movie S5 (.avi format). Diffusion, dissociation, and reassociation of an individual ternary complex in the MSK meshwork, as obtained from spatial simulations.
  
• movie S6 (.avi format). Experimentally observed receptor rebinding events.
  
• movie S7 (.avi format). Transient receptor confinement at the native cell PM observed by fast TALM.